Polymerase chain reaction (PCR) is one of the common techniques in molecular biology, which can amplify nucleic acids through the cycle of denaturation, annealing and extension. Based on the principle of common PCR, rapid PCR is to realize the amplification of nucleic acids in less time without affecting the specificity, sensitivity and fidelity of the reaction. A lot of research work in this field has been going on in recent years. This article will make a review of the development of rapid PCR with emphases on the improvement of DNA polymerase, the choice of additives and the improvement of thermocyclers.

Previous studies have revealed Twist,a basic helix-loop-helix transcription factor,plays an important role in breast cancer metastasis.To clarify the molecular mechanism of its involvement in cancer metastasis,Twist was silenced by RNAi in highly metastatic 4T1 cells.Then microarray chips were used to investigate the gene-expression pattern of the Twist-knockdown 4T1 cells and the normal 4T1 cells.The results indicated that silencing of Twist significantly suppressesed lung metastasis of 4T1 cells in vivo.Direct comparison of gene-expression profiles showed that 167 genes in Twist-knockdown cells differed dramatically in expression levels from those in control cells.Among the 167 genes,26 well-known tumor-associated genes,including 15 up-regulated and 11 down-regulated genes were found.These genes appear to be regulated by Twist during breast tumorigenesis.The findings provide new insights into the mechanism by which Twist is involved in tumorigenesis.

OBJECTIVE: To evaluate the status of application of continuous renal replacement therapy (CRRT) in patients in the intensive care unit (ICU). METHODS: The clinical data of 171 ICU patients who had indications for CRRT in the First Affiliated Hospital of Xi'an Jiaotong University from January to December 2014 were retrospective analyzed. The patients were divided into non-CRRT group (n=23) and CRRT group (n=148) which was composed of continuous veno-venous hemofiltration (CVVH) group (n=66) and non-CVVH group (n=82). Among the 148 CRRT patients, 83 had improvement, 46 had no improvement, and 19 died. Among the 66 CVVH patients, 39 had improvement, and 27 had no improvement. The indications of the 171 patients and several aspects of the CRRT group including anticoagulation regimen, catheterization program, replacement fluid, and CRRT pipe flushing were statistically analyzed. And the correlations between the clinical data, CRRT mode, and efficacy were evaluated. RESULTS: The rate of rational indications of 171 patients was 86.55%, meanwhile the rates of the reasonability of anticoagulation method, catheterization program, replacement fluid, and CRRT pipe flushing of the CRRT group were 84.76%, 66.89%, 100%, and 100%, respectively. Single factor analysis showed that the levels of BUN (P=0.035) and creatinine (P=0.007) at baseline and the urine output on the third day (P=0.034) had statistically significant difference among the CVVH group, non-CVVH group, and non-CRRT group. There were also significant differences in the survival rate (P=0.01) and prognosis of survivals (P=0.007) in CRRT group among different catheterization programs. Multivariate analysis indicated that there was significant correlation between the lengh of stay in ICU [P=0.005, OR: 0.734, 95% CI(0. 592, 0.910)], catheterization program [P=0.016, OR: 5.302, 95%CI(1.369, 20.527)] and prognosis. CONCLUSION: The indications, anticoagulation method, and catheterization program of the CRRT group were partially unreasonable in the 171 patients in our study, and the unreasonability rates were 13.45%, 15.24%, and 33.11%, respectively. Clinical pharmacists should focus on the drug regimens and individual monitoring of ICU patients in the process of CRRT, considering their pathophysiological characteristics and complexity of medical therapeutic schemes.

AIM: Recent studies showed that stem cells could replace injured cardiomyocyte and increase the number of functional cardiomyocytes. Researching the pertinent literature in China Journal Full-text Database (CJFD) published between 2005 and 2008 indicated that the researches on stem cell transplantation in the treatment of primary dilated cardiomyopathy were few. This study investigated the therapeutic efficacy and security of peripheral blood stem cell transplantation in coronary artery in treatment of dilated cardiomyopathy and its effects on left ventricular function. METHODS: Forty-two patients with dilated cardiomyopathy between December 2006 and September 2007 were enrolled at the Department of Cardiology of First People's Hospital of Yunnan, including twenty-eight males and fourteen females, averagely aged (56?3) years. Inclusive criteria: patients with less than 65 years, left ventricular enlargement, and left ventricular ejection fraction (LVEF) ≤ 45%, and without coronary artery disease after coronary arteriongraphy. Informed consents were obtained from patients. Patients were divided into stem cell transplantation group (n=15) and control group (n=27) on the basis of whether being treated by stem cell transplantation. Patients in the stem cell transplantation group were consecutively administered granulocyte colony-stimulating factor (G-CSF) by hypodermic injection to stimulate bone marrow stem cells themselves based on conventional treatment for five days. Peripheral blood stem cell suspension was disassociated on the 6th day, and the collected suspension was injected into left anterior descending branch over the wire saccule tube for autologous peripheral blood stem cell transplantation. Patients in the control group were administered by conventional treatment of dilated cardiomyopathy. Security and adverse reaction were observed during the mobilization, collection and returning injection of peripheral blood stem cell by coronary artery. Morphous, cardiac function and motion index of left ventricle wall were evaluated using ultrasoundcardiogram before and 3 months after transplantation. Survival rate and incidence rate of heart incidents were compared. RESULTS: Three months after stem cell transplantation in coronary artery, there were a significant decrease in cardiac end-systolic volume (ESV), cardiac end-diastolic volume (EDV) and motion index of left ventricle wall, but a significant increase in LVEF(P

Embryonic stem cells(ESCs) are pluripotent cells derived from the inner cell mass of blastocyst-stage embryos, possessing permanent self-renewal and indefinite proliferative capacity in vitro. And ESCs could differentiate into the hematopoietic cell fate. Therefore ESCs may provide an alternative for hematopoietic stem cell transplantation and blood cells transfusion. Furthermore, ESCs differentiation could also provide a powerful model system to better understand the hematopoietic development and the mechanism involved. The current status for efforts to differentiate ESCs into hematopoietic lineages were reviewed.

OBJECTIVETo observe the clinical efficacy difference in treatment of functional dyspepsia (FD) between syndrome differentiation based acupuncture and ordinary acupuncture.

METHODSSeventy FD patients were assigned to a syndrome differentiation based acupuncture group (Group A) and an ordinary acupuncture group (Group B) by Excel Software randomization. Zhongwan (RN12 ), Tianshu (ST25), and Zusanli (ST36) were needled as main points for patients in Group A. Meanwhile, different combined acupoints were needled according to syndrome differentiation. Only the same main points were needled for patients in Group B. All patients were needled once per day, 30 min each time, 6 days as one treatment cycle, 2 treatment cycles in total. Fasting serum levels of gastrin (GAS) and motilin (MTL) were determined before treatment and after 2 treatment cycles. 36-item Short-form Heath Survey (SF-36) and Nepean Dyspepsia Index [NDI, including Nepean Dyspepsia Symptom Index (NDSI) and Nepean Dyspepsia Life Quality Index (NDLQI)] were assessed before treatment, after 2 treatment cycles, and one month after treatment.

RESULTSCompared with before treatment in the same group, serum levels of GAS and MLT increased in the two groups after 2 treatment cycles (P <0. 05), but changes were more obvious in Group A (P <0. 05). Compared with before treatment in the same group, SF-36 and NDLQI score increased, and NDSI score decreased in the two groups after 2 treatment cycles and 1 month after treatment (all P <0. 05). Compared with Group B, SF-36 and NDLQI score increased in Group A after 2 treatment cycles and 1 month after treatment (P <0. 05, P <0. 01). But NDSI score at 1 month after treatment was lower in Group A than in Group B (P <0.01).

CONCLUSIONSyndrome differentiation based acupuncture could evidently improve dyspeptic symptoms of FD patients, and significantly improve their quality of life with remarkable curative effect.

Acupuncture Points ; Acupuncture Therapy ; Dyspepsia ; therapy ; Humans ; Motilin ; Needles ; Quality of Life ; Surveys and Questionnaires ; Syndrome

OBJECTIVETo investigate the protective effect of Danxuetong injection (DXT, a combination of Danshen and Xueshuantong injections) against testicular ischemia-reperfusion injury following testis torsion/detorsion in rats.

METHODSThirty-two 4-week-old healthy male SD rats were randomly divided into four groups of equal number: sham operation, normal saline, single DXT injection, and successive DXT injection. The rat models of testicular ischemia-reperfusion injury were established by 2-hour 720-degree torsion/detorsion of the unilateral testis. At 6 weeks after modeling, the rats were killed and their testes were harvested for measure- ment of testicular coefficients, sperm counts, sperm motility, and the levels of total anti-oxidative capacity (T-AOC) , superoxide dismutase (SOD) , nitric oxide synthase (NOS) , and malondialdehyde ( MDA) in the testis tissue.

RESULTSCompared with the rats of the normal saline group, those of the single DXT injection and successive DXT injection groups showed significant increases in the testicular coefficient (0.11 ± 0.03 vs 0.35 ± 0.04 and 0.40 ± 0.06, P < 0.05), sperm count ([0.46 ± 0.10] vs [1.44 ± 0.50] and [3.00 ± 1.28] x10(9)/ml, P < 0.05), sperm motility ([13.63 ± 14.04] vs [39.63 ± 5.04] and [76.31 ± 3.67]%, P < 0.05), the activity of SOD (72.76 ± 5.58 vs 116.25 ± 8.83 and 133.20 ± 13.84, P < 0.05), and the level of T-AOC (5.58 ± 1.07 vs 13.34 ± 5.81 and 19.21 ± 5.69, P < 0.05), but a remarkable decrease in the content of MDA (42.38 ± 8.94 vs 20.94 ± 5.65 and 15.02 ± 1.03, P < 0. 05) in the injured testes.

CONCLUSIONDXT can effectively rid the testis tissue of oxygen free radicals, improve sperm count and motility by antioxidation, and protect the testis tissue of prepubertal rats against testicular ischemia-reperfusion injury after testis torsion/detorsion. It also has a protective effect on the contralateral testis, and successive injection has a better effect than single injection of DXT.

Animals ; Antioxidants ; therapeutic use ; Drug Therapy, Combination ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Spermatic Cord Torsion ; complications ; therapy ; Superoxide Dismutase ; metabolism ; Testis ; blood supply ; metabolism

OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome

OBJECTIVETo evaluate the impact of nicotine- and tar-free cigarette smoke extract (fCSE) on the serum testosterone (T) level and erectile function of male rats.

METHODSWe randomized 30 male SD rats to three groups of equal number to receive subcutaneous injection of PBS (1.0 ml / 300 g body weight per day), fCSE (1.0 ml/300 g body weight per day), and reduced glutathione hormone (GSH, 200 mg per kg body weight per day) in addition to fCSE (fCSE + GSH), respectively, all for 8 weeks. Then we evaluated the erectile function of the rats by measuring the maximal intracavernous pressure (MICP), mean arterial pressure (MAP), ICP/MAP ratio, time of stimulation to MICP (Tmax), and cavernosal filling fate (CFR). We determined the serum T level, the activities of superoxide dismutase (SOD) , malondialdehyde (MDA), and nitric oxide synthase (NOS) in the cavernosal tissue, and also observed the morphological changes of the corpus cavernosum.

RESULTSCompared with the controls, the rats of the fCSE group showed obvious decreases in the levels of serum T ([5.37 ± 1.43] vs [3.22 ± 1.11] μg/L), NOS ([2.90 ± 0.27] vs [1.67 ± 0.18] U/mg) , and SOD ([18.41 ± 1.09] vs [13.36 ± 1.18] U/mg prot) and erectile function-related indexes MICP ([85.92 ± 6.36] vs [58.99 ± 10.76] mmHg), MICP/MAP (0.86 ± 0.09 vs [0.56 ± 0.08]), and CFR (2.14 ± 0.44 vs 0.89 ± 0.44), but markedly increased Tmax ([29.90 ± 5.78] vs [42.90 ± 8.56]s), with a positive correlation between the serum T level and CFR (r = 0. 364, P < 0.05). Masson staining revealed a lower ratio of the corpus cavernosum smooth muscle tissue to collagen fiber in the fCSE group (0.27 ± 0.04) than in the control (0.98 ± 0.12). Compared with the fCSE group, the fCSE + GSH group exhibited significantly improved MICP ([58.99 ± 10.76 ] vs [77.95 ± 7.71] mmHg), MICP/MAP (0.56 ± 0.08 vs 0.77 ± 0.09), and CFR (0.89 ± 0.44] vs 1.76 ± 0.42) and shortened Tmax ([42.90 ± 8.56 ] vs [32.10 ± 5.84 ] s). The ratio of the corpus cavernosum smooth muscle tissue to collagen fiber was higher in the fCSE + GSH than in the fCSE group (0.77 ± 0.09 vs 0.27 ± 0.04) but still lower than in the control (0.98 ± 0.12).

CONCLUSIONNicotine- and tar-free cigarette smoke extract reduces the serum T level and erectile function of rats, which is related to oxidative stress. Antioxidant therapy can improve erectile function but has a limited value for morphological protection of the penile tissue.

Animals ; Erectile Dysfunction ; chemically induced ; Male ; Malondialdehyde ; metabolism ; Muscle, Smooth ; pathology ; Nicotine ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Superoxide Dismutase ; metabolism ; Tars ; Tobacco ; adverse effects

Objective To explore the clinical value of renal dynamic imaging and urinary N-acetyl-?-D-glucosaminidase(NAG),apoptosis DNA fragment(ADF) in evaluating the damage degree of hydronephrotic kidneys(HnK) in children with hydronephrosis.Methods Level of glomerular filtration rate(GFR) was detected in 41 children with congenital hydronephrosis by renal dynamic imaging,and urine NAG,ADF in pelvis in HnK and healthy kidneys (as controls) were detected by enzyme-linked immuno-sorbent assay(ELISA).Patholo-gic changes of HnK in 41 children were graded intoⅠ~Ⅴ according to Elder standard. And GFR,urinary NAG and ADF of HnK were divi-ded into subgroup according to pathologic changes ,at the same time statistical analysis was performed within each groups. And the correlations of pathologic grades with GFR,urinary NAG and ADF of HnK were analyzed.Results 1.Kindneys GFR in healthy kidneys and Hnk were (174.33?20.43)?10-3 L/min,(143.86?17.51)?10-3 L/min respectinely,and there was significant difference between healthy kidneys and Hnk (P0.05).3.There was significant negative correlation between GFR levels of HnK and pathologic grades(r=-0.814 P0.05).Conclusions For hydronephrotic kidneys,urinary NAG can eva-luate impaired nephric tubule whereas renal dynamic imaging may evaluate the damage level of glomeruli;urine ADF may not indicate the damage level of diseased kidneys in children with congenital hydronephrosis.