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Candida is an intractable life-threatening pathogen. Candida infection is extremely difficult to eradicate, and thus is the major cause of morbidity and mortality in immunocompromised individuals. Morevover, the rapid spread of drug-resistant fungi has led to significant decreases in the therapeutic effects of clinical drugs. New anti-Candida agents are urgently needed to solve the complicated medical problem. Natural products with intricate structures have attracted great attention of researchers who make every endeavor to discover leading compounds for antifungal agents. Their novel mechanisms and diverse modes of action expand the variety of fungistatic agents and reduce the emergence of drug resistance. In recent decades, considerable effort has been devoted to finding unique antifungal agents from nature and revealing their unusual mechanisms, which results in important progress on the development of new antifungals, such as the novel cell wall inhibitors YW3548 and SCY-078 which are being tested in clinical trials. This review will present a brief summary on the landscape of anti-Candida natural products within the last decade. We will also discuss in-depth the research progress on diverse natural fungistatic agents along with their novel mechanisms.
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Humans , Antifungal Agents/pharmacology , Biological Products/pharmacology , Candida/drug effects , Candidiasis/drug therapy , Microbial Sensitivity TestsABSTRACT
Objective:To observe the efficacy of apatinib combined with capecitabine in the treatment of advanced esophageal cancer.Methods:A total of 101 patients with advanced esophageal cancer in Taixing People's Hospital of Jiangsu Province from June 2017 to February 2018 were enrolled, and all the patients were divided into the control group (50 cases) and the observation group (51 cases) according to the random number table. The control group was treated with capecitabine combined with radiotherapy, and the observation group was treated with apatinib on the basis of the control group. The therapeutic effects, adverse reactions and progression-free survival (PFS) time of the two groups were compared.Results:The overall response rate in the observation group was higher than that in the control group [90.2% (46/51) vs. 72.0% (36/50)], and the difference was statistically significant ( χ2 = 5.473, P = 0.019). There were no significant differences in leukopenia, neutropenia, thrombocytopenia, anemia, proteinuria and hypertension between the two groups (all P > 0.05). The median PFS time in the observation group was 18.49 months (95% CI 15.35-25.03 months), and that in the control group was 13.33 months (95% CI 10.36-18.24 months), and the difference between the two groups was statistically significant ( χ2 = 5.995, P < 0.01). Conclusions:The therapeutic effect of apatinib combined with capecitabine in the treatment of advanced esophageal cancer is accurate. No obvious adverse reaction occurs, and the PFS time is prolonged.
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The proportion of laparoscopic surgeries is continuously increasing in general surgeries. Along with the development and application, new image sensor and digital image processing technology accelerated the emergence of novel laparoscope in recent years. Stereoscopic laparoscope (3D) appearing make the space orientation more accurate. new imaging methods and new structure design satisfy more clinical requirements; combination with optical technology (NBI technique, PDD technique, ICG technique) make intraoperative diagnosis possible.
Subject(s)
Image Processing, Computer-Assisted , Laparoscopes , LaparoscopyABSTRACT
Drosophila neural development undergoes extensive chromatin remodeling and precise epigenetic regulation. However, the roles of chromatin remodeling in establishment and maintenance of cell identity during cell fate transition remain enigmatic. Here, we compared the changes in gene expression, as well as the dynamics of nucleosome positioning and key histone modifications between the four major neural cell types during Drosophila neural development. We find that the neural progenitors can be separated from the terminally differentiated cells based on their gene expression profiles, whereas nucleosome distribution in the flanking regions of transcription start sites fails to identify the relationships between the progenitors and the differentiated cells. H3K27me3 signal in promoters and enhancers can not only distinguish the progenitors from the differentiated cells but also identify the differentiation path of the neural stem cells (NSCs) to the intermediate progenitor cells to the glial cells. In contrast, H3K9ac signal fails to identify the differentiation path, although it activates distinct sets of genes with neuron-specific and glia-related functions during the differentiation of the NSCs into neurons and glia, respectively. Together, our study provides novel insights into the crucial roles of chromatin remodeling in determining cell type during Drosophila neural development.
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Objective To evaluate the effects of the size of lymph node metastasis (LNM) on the chemoradiotherapy efficacy and prognosis for the patients after resection of thoracic esophageal squamous cell carcinoma (ESCC).Methods Between 2011 and 2014,a total of 75 esophageal squamous carcinoma patients with secondary LNM after resection of ESCC were recruited in this retrospective study.They were treated with curative radiotherapy only or concurrent chemoradiotherapy in the Affiliated Taixing People's Hospital of Yangzhou University.Thc LNM volume and maximum diameters were measured by the Monaco treatment planning system.The enrolled patients were grouped according to the median values of LNM volume and maximum diameters.The relationship between the responsiveness to treatment and these markers was analyzed by univariate and multivariate logistic analysis.The Kaplan-Meier method and Log-rank test were adopted to calculate and compare the overall survival (OS) rates with these markers.The Cox proportional hazards model was used to carry out univariate and multivariate analyses.Results The overall effective rate was 69.3％ for all enrolled patients.The response rates were 81.6％ with LNM volume ＜57 cm3 and 56.8％ with LNM volume ≥57 cm3.The response rates were 83.8％ with LNM maximum diameter ＜ 5 cm and 55.3％ with LNM maximum diameter ≥5 cm.The responses to treatment were highly associated with treatment method (OR =1.825,95％ CI:1.134-3.658,P =0.017),LNM volume (OR =4.183,95％ CI:1.416-12.354,P =0.010) and maximum diameter (OR =3.374,95％ CI:1.185-9.611,P =0.023) by univariate logistic regression analysis.Multivariate logistic regression analysis showed that therapeutic method (OR =1.225,95％ CI:1.085-2.837,P =0.038) and LNM volume (OR =1.614,95％ CI:1.003-3.025,P =0.048) were independent risk factors for tumor response.The median OS time of this cohort was 14 months,and the 1,2 and 3 year OS rates were 60.7％,25.3％ and 20.1％,respectively.Kaplan-Meier survival analysis revealed that TNM stage (HR =2.039,95 ％ CI:1.234-3.370,P =0.005),treatment methods (HR =1.858,95 ％ CI:1.385-2.958,P =0.013),LNM volume (HR =2.642,95％ CI:1.552-4.497,P ＜ 0.001) and LNM maximum diameter (HR =3.399,95 ％ CI:1.939-5.958,P ＜ 0.001) were significantly associated with OS.Furthermore,multivariate Cox proportional hazard regression model analysis for OS was performed and the results showed that TNM stage (HR =2.023,95 ％ CI:1.149-3.560,P =0.015),LNM volume (HR =2.055,95 ％ CI:1.041-4.055,P =0.038) and maximum diameter (HR =1.910,95％ CI:1.137-3.895,P =0.045) were considered as independent prognostic risk factors for OS.Conclusion LNM volume in ESCC patients with secondary LNM after esophagectomy has great values for predictive therapeutic effects and survival outcomes,and LNM maximum diameter has significant value for survival outcomes.
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OBJECTIVE@#To obtain three-dimensional intraosseous artery of the hamate and to provide the vascular anatomy basis of hamate fracture fixation.@*METHODS@#PbO (lead monoxide, Sinopharm Chemical Reagent Beijing Co. Ltd) was ground into particles less than 40 μm and suspended in turpentine oil (Chemical Reagent Beijing Co. Ltd) at ratios of 1 g : 1.5 mL, 1 g : 1 mL and 1 g : 0.5 mL. Three specimens were investigated. Brachial arteries were cannulated and perfused with lead-based contrast agent. Hamates were harvested and scanned using micro-computed tomography (microCT). The acquisition protocols were as follows: CT scan setup: total rotation [Degrees], 360; rotation steps, 360; X-ray detector setup: transaxial, 2048; axial, 2048; exposure time, 1 500 ms, Binning, 1; system magnification: high-med. X-ray tube setup: 80 kV, 500 mA current. The down-sampling factor used in the reconstruction was 2. The effective voxel size of the final image was 27.30 μm. The three-dimensional model of the hamate was generated and the distribution and pattern of vessels were evaluated.@*RESULTS@#There were abundant extraosseous vessels around the hamate. They were mainly running in the tendons and ligaments around the hamate. Four vascular zones were identified on the hamate surface. They were on the palmar platform of the hamate body, on the dorsal side, on the ulnar side and on the tip of hamulus, namely. There were anastomoses among 4 vascular zones. We did not observe any vessels penetrating through the articular cartilage. The extraosseous vessels of the vascular zones gave a number of intraosseous branches into the hamate. The hamate body received intraosseous blood supply from the dorsal, palmar and ulnar while the hamulus from the palmar, ulnar and hamulus tip. There were some intraosseous branches anastomosing with each other.@*CONCLUSION@#The extraosseous and intraosseous vessels of the hamate were more than what used to be considered. The hamate body and hamulus received blood supply from multiple directions and arteries anastomosed extensively both outside and inside the hamate, making it possible that the intraosseous perfusion survived after fracture. It is likely that the nonunion after the hamate fracture is not caused by the vascular damage but the malalignment of the fragments.
Subject(s)
Humans , Beijing , Brachial Artery , Fluoroscopy , Fractures, Bone/diagnostic imaging , Hamate Bone/injuries , Ulna , Wrist Injuries/diagnostic imaging , X-Ray MicrotomographyABSTRACT
Objective@#To apply Nutritional Risk Screening-2002(NRS-2002) to perform nutritional status score for the patients with thoracic esophageal squamous cell carcinoma (ESCC) receiving surgery, and to explore the prognostic impact of long-term survival.@*Methods@#A total of 117 patients who were diagnosed with ESCC from 2010 to 2012 were retrospectively analyzed. They recieved standard curative esophagectomy in the Yangzhou University Affiliated Taixing People′s Hospital. The nutritional status and risk score for recruited patients were assessed according to the standard of NRS-2002 tool prior to surgery, and these patients were grouped for further analysis according to the median values of NRS-2002 score. Finally, the relationship between NRS-2002 score and prognosis was analyzed.@*Results@#Patients were classified into two groups, with 45 in the NRS-2002<2.0 group, and 72 cases in the NRS-2002≥2.0, respectively. In the NRS-2002<2.0 group, the 1-, 3-, and 5-year progression-free survival (PFS) rates were 75.6%, 44.4% and 40.0% separately, while in the NRS-2002≥2.0 group, the PFS rates were 61.1%, 6.9% and 4.2% respectively, and the differences were statistically significant (P<0.001). Correspondingly, in the NRS-2002< 2.0 group, the 1-, 3-, and 5-year overall survival (OS) rates were 97.8%, 66.7% and 57.8% separately, while in the NRS-2002≥2.0 group, the OS rates were 91.7%, 33.3% and 16.7% respectively, and the differences were also statistically significant (P<0.001). Univariate analysis showed that N stage, TNM stage and NRS-2002 score were closely related to PFS and OS (P<0.05), and T stage was only associated with OS in patients with thoracic esophageal squamous cell carcinoma (P<0.05). Furthermore, multivariate Cox regression analysis showed that N stage (RR=1.640, 95% CI 1.049-2.565, P=0.030) and NRS-2002 (RR=3.154, 95% CI 1.946-5.113, P<0.001) were independent prognostic factors for PFS in patients with ESCC after surgery. Additionally, pathological differentiation (RR=1.556, 95% CI 1.004-2.440, P=0.041), N stage (RR=1.624, 95% CI 1.017-2.593, P=0.042) and NRS-2002 (RR=3.906, 95% CI2.245-6.795, P<0.001) were independent prognostic factors for OS in ESCC patients following surgery.@*Conclusion@#Preoperative nutritional risk screening NRS-2002 score is an independent prognostic factor in patients with ESCC receiving surgery and could be used as a tool for primary screening for nutritional risk.
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Objective To investigate the effect of pidotimod granules and Carboxymethyl starch sodium solution in treating children with recurrent respiratory tract infection curative effect and effect on immune function of children.Methods 128 cases of children with recurrent respiratory tract infection in our hospital from June 2013 to June 2016 were selected and divided into treatment group and control group with 64 cases in each group. Children in the control group were treated with Carboxymethyl starch sodium solution, the treatment group was given pidotimod granules on the basis of the control group.Two groups of children were treated for 12 weeks.The therapeutic effects of the two groups were compared, the changes of immune function before and after treatment, cough, fever, lung rales disappear time, and the incidence of adverse reactions.Results The total effective rate of the treatment group (90.62%) was higher than the control group (75.00%), the difference was statistically significant (P<0.05), the levels of IgA, IgM and IgG increased after treatment in both groups, the levels of IgA, IgM and IgG in the treatment group were higher than those in the control group, the difference was statistically significant (P<0.05), the levels of CD3 +, CD4 +and CD4 +/CD8 +increased after treatment in both groups, and the levels of CD3 +, CD4 +and CD4 +/CD8 +in the treatment group were higher than those in the control group, the difference was statistically significant (P<0.05), the treatment group in children with cough, fever, pulmonary rales disappeared faster than the control group, the difference was statistically significant (P <0.05), two groups were no obvious adverse reactions.Conclusion Pidotimod granules combined with carboxymethyl starch sodium solution in treating children with recurrent respiratory tract infections is effective, and could significantly improve the immune function of children.
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The paper takes literature in the Health Technology Assessment (HTA) field in Web of Science and " China Academic Journal Network Publishing Database" as its study subject,draws scientific knowledge map with EXCEL,CiteSpace on data like high-frequency keywords and burst terms and analyzes study hotspots and evolution trend.It suggests that HTA theoretical system be perfected,study be converted into demonstration and study cooperation be strengthened to promote study and development of China's HTA.
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Objective To investigate the effects of fruit potassium supplement on hypokalemia at in the perioperative period of percutanous coronary intervention(PCI)? Methods Ninety cronary herat disease patients with hypokalemia undergoing PCI during July to December 2011 involved the study? Since admission,the patients were administered with oranges and bananas each 500 g daily until the third day after PCI,on the basis of oral administration of potassium? Serum potassium was assessed in all patients at time points of the day at PCI,the first day and third day after PCI? Results The time range for the patients to take potassium supplement by eating fruits was 12-45 hours,averaged(28?84 ± 11?70)hours? The serum potassium levels on days 1 and 3 were significantly higher than that on the day for the admission(P < 0?05 for both)and even maintained at the normal level? Conclusion The oral complement of potassium by eating bananas and oranges can help patients to correct and prevent hypokalemia during the perioperative period of PCI?
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Sustained activation of sympathetic nervous system in response to stimulation of a wide variety of stress factors is an independent risk factor for the development of essential hypertension. Adrenal hormone biosynthesis pathway as an important part of the sympathetic nervous system consists of hormones, neurotransmitters, receptors, and a variety of synthases and invertases. In this article, we have systematically reviewed research progresses made in elucidating the interactions between genes of the adrenal hormone biosynthesis pathway and stress factors in the pathogenesis of essential hypertension.
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Animals , Humans , Hormones , Metabolism , Hypertension , Genetics , Metabolism , Pathology , Sympathetic Nervous System , Metabolism , PathologyABSTRACT
Due to an increased risk of infection, dental implant in organ transplantation patients has long been considered questionable, particularly when the restoration is complicated. Five-year follow-up data of a 45-year-old liver transplant recipient with long-term immunosuppressive therapy was reported. One year after liver transplantation, 11 Brånemark implants were inserted in the maxilla and mandible, using minimally invasive surgery. Oral clinical parameters included peri-implant bone absorption, probing depth, and implant mobility. The measured fifth-year parameters were within normal ranges indicating a stable osseointegration with moderate vertical bone loss. This case report suggests that immunocompromised patients can be successfully rehabilitated with dental implants through careful examination, suitable antibiotic administration, and minimally invasive dental implant procedure.
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Humans , Male , Middle Aged , Dental Implants , Immunosuppressive Agents , Therapeutic Uses , Liver Transplantation , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To elucidate the development of mapping and localization of susceptible genes on chromosomes to asthma related phenotypes.</p><p><b>DATA SOURCES</b>Published articles about susceptibility genes for asthma related phenotypes were selected using PubMed.</p><p><b>STUDY SELECTION</b>Using methods of candidate gene positional clone and genome-wide scan with linkage and association analysis to determine the location in the genome of susceptibility genes to asthma and asthma related phenotypes.</p><p><b>RESULTS</b>There are multiple regions in the genome harboring susceptibility genes to asthma and asthma related phenotypes, including chromosomes 5, 11, 12, 6, 2, 3, 13, 7, 14, 9, 19 and 17. Many of these regions contain candidate genes involved in asthma development and progression. Some susceptible genes may affect the phenotype expression or response to therapy. In addition, the interaction of multiple genes with the environment may contribute to the susceptibility to asthma.</p><p><b>CONCLUSIONS</b>As an essential step toward cloning the susceptible genes to asthma, fine mapping and localization on chromosomes are definitely needed. Novel powerful tools for gene discovery and the integration of genetics, biology and bioinformatics should be pursued.</p>
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Humans , Asthma , Genetics , Chromosome Mapping , Methods , Genetic Predisposition to Disease , GeneticsABSTRACT
Objective To investigate the observation and nursing for patients with atria] fibrillation (AF)after left atrial linear ablation surrounding ipeilateral pulmonary veins (PVs). Methods Thirty patients with persistent AF underwent left atrial linear ablation surrounding ipsilateral PVs guided by CARTO electroanatomic mapping system and double Lasso catheters technique. The end point of ablation was defined as absence of all PV spikes after isolation and bidirectional conduction block between left atrial and PVs. Results After a median of (245±65)days of follow-up, twenty-one patients were free of AF. Eight patients subjected recurrent atrial tachyarrhymias, including 5 typical AFL, 2 atrial taehycardia and 1 paroxysmal AF. Two patients with persistent AF remaining PV spikes in left superior PV failed to be translated to sinus rhythm. Nine patients underwent repeated ablation and 8 of them were free of atrial taehyarrhythmia after (192±92) days follow-up. Total success rate after two procedures was 92.8%. Conclusions Nurses should perform effective care according to the symptom of patients before, during and after ablation procedure and relieve the patients from pain.
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<p><b>OBJECTIVE</b>To construct the haplogroup and perform an analysis of mitochondrial whole-genome sequence in Tibetan and Han Chinese. Variations of nucleotide of mitochondrial DNA (mtDNA) were identified and compared between the Tibetan and Han population.</p><p><b>METHODS</b>The mtDNA whole sequences of 40 Tibetan and 50 Han individuals were sequenced by an Applied Biosystems 3730 DNA automatic sequencer. The sequences were assembled using software phredPhrap16.0, and all assembled sequences were manually verified according to the criterion of rCRS (revised Cambridge Reference Sequence). The haplogroups of mtDNA were constructed using phylogenetic analysis according to the criteria of MITOMAP by Network method. The data were elucidated by integrated methods.</p><p><b>RESULTS</b>Authors' results showed that all the pooled 90 subjects belonged to the Macrohaplogroup M and N, and were classified into 13 haplogroups. No differences were observed among the haplogroups of the two populations except for M9 haplogroup. A total of 21 variants were detected by comparing the mtDNA whole sequences between Tibetan and Han population; of those, 5 variants have not been reported before. In addition, we constructed the haplotypes of 5 variants harboring the D-loop region, and founded prominent difference in both supertype 1 and supertype 2 between Tibetan and Han population.</p><p><b>CONCLUSION</b>The phylogenetic analysis indicates that the Tibetan and Han ethnic groups shared close maternal relationship in origin. The biological implication of the significant variants is worth elucidating; whether they are the results of adaptive selection or neutral selection or pathological variations need to be further studied.</p>
Subject(s)
Humans , Asian People , Genetics , China , Ethnology , DNA, Mitochondrial , Ethnicity , Genetics , Evolution, Molecular , Genetics, Population , Genome, Mitochondrial , Genetics , Haplotypes , Genetics , Tibet , EthnologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the influence of Tinglizi on collagen volume fraction (CVF) and perivascular collagen volume area (PVCA ) in left ventricle tissue of cardiac hypertrophy induced by abdominal aortic banding in rats.</p><p><b>METHOD</b>Ventricular remodeling was induced by abdominal aortic banding (AAB) in rats. After 30 day treatment, the systolic blood pressure (SBP), diastolic blood pressure (DBP); heart rate (HR) were measured. The histological assay consisted of the HE stain for determining the myo-cardium cell cross section and collagen stain (Van Gieson' method) for determining collagen content, including collagen volume fracton (CVF) and perivascular collagen volume area (PVCA).</p><p><b>RESULT</b>The experimental data demonstrated that Tinglizi decreased SBP, DBP, HR and could significantly reduce the total collagen content (CVF, PVCA) and lessen the myocardium cell cross section (P < 0.05).</p><p><b>CONCLUSION</b>Tinglizi may decrease the total collagen content of ventricle and attenuate the ventricular remodeling induced by abdominal aortic banding.</p>
Subject(s)
Animals , Male , Rats , Blood Pressure , Cardiomegaly , Drug Therapy , Drugs, Chinese Herbal , Pharmacology , Heart Rate , Heart Ventricles , Rats, Sprague-Dawley , Ventricular RemodelingABSTRACT
<p><b>OBJECTIVE</b>To evaluate the safety and efficacy of sublingual immunotherapy with 'Dermatophagoides Farinae Drops' in D. farinae allergic asthma and/or rhinitis patients.</p><p><b>METHODS</b>A 25-week double-blind, placebo-controlled, multi-centered trail was conducted in 278 children (aged 4 - 18 yr) with mite-induced asthma and/or rhinitis. Patients were randomly assigned to receive sublingual immunotherapy (SLIT) with 'Dermatophagoides Farinae Drops' (n = 139) or placebo (n = 139) for 25 weeks and the dosage and administration strictly followed the manufacturer's instructions. At the beginning of the 2nd, 3rd, 4th, 6th, 10th, 14th, 18th, 22nd week of the treatment, the patients were asked to accept follow-up visit, during the clinical trial all patients and parents were asked to keep a daily record of their asthma symptom scores, rescue medicine use, rhinitis symptom scores, morning and evening peak expiratory flow. Asthma symptom scores, reduction in use of rescue medicine, rhinitis symptom scores, lung function tests, skin sensitivity to mite, mite-specific immunoglobulin (Ig) E and IgG4, and quality of life and adverse effect were assessed during the study.</p><p><b>RESULT</b>(1) Of the 278 children, 27 dropped out before the study completion. (2) After 25 weeks of treatment, the median variability of PEFR was -1.38 for SLIT group and -0.90 for the placebo (P < 0.05). (3) Besides, the mean variability of medicine score of asthma was -0.08 for SLIT group and 0.52 for the plcebo (P < 0.05). (4) The median variability of rhinitis symptom score was -1.96 for SLIT group and -1.03 for the placebo (P < 0.01). (5) The rescue medicine usage of SLIT reduced but did not show significant differences between SLIT and placebo. (6) After 25 weeks treatment, the increase of D. farinae specific IgE antibody of two groups were similar, while specific IgG4 increased significantly in SLIT compared to the patients in control one (P < 0.01); (7) No severe adverse events happened in the trial and the most-likely adverse events were mild asthma and local rash.</p><p><b>CONCLUSION</b>Dermatophagoides Farinae Drops is safe and effective in treating allergic asthma and atopic rhinitis.</p>
Subject(s)
Adolescent , Animals , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Asthma , Therapeutics , Dermatophagoides farinae , Chemistry , Metabolism , Double-Blind Method , Mites , Chemistry , Rhinitis , TherapeuticsABSTRACT
Human genome has structures of haplotype and haplotype block which provide valuable information on human evolutionary history and may lead to the development of more efficient strategies to identify genetic variants that increase susceptibility to complex diseases. Haplotype block can be divided into discrete blocks of limited haplotype diversity. In each block, a small fraction of ptag SNPsq can be used to distinguish a large fraction of the haplotypes. These tag SNPs can be potentially useful for construction of haplotype and haplotype block, and association studies in complex diseases. There are two general classes of methods to construct haplotype and haplotype blocks based on genotypes on large pedigrees and statistical algorithms respectively. The author evaluate several construction methods to assess the power of different association tests with a variety of disease models and block-partitioning criteria. The advantages, limitations and applications of each method and the application in the association studies are discussed equitably. With the completion of the HapMap and development of statistical algorithms for addressing haplotype reconstruction, ideas of construction of haplotype based on combination of mathematics, physics, and computer science etc will have profound impacts on population genetics, location and cloning for susceptible genes in complex diseases, and related domain with life science etc.
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Humans , Algorithms , Computational Biology , Computer Simulation , Haplotypes , Genetics , Mathematics , Methods , Models, Genetic , Polymorphism, Single Nucleotide , GeneticsABSTRACT
Human tumstatin(hTumstatin)cDNA was amplified from recombinant plasmid pET-3c-tum, cloned in frame with the signal sequence in yeast vector pPICZalphaA and transformed into Pichia pastoris GS115 by electroporation. The expression of hTumstatin in GS115(pPICZalpha-tum)was then induced by methanol and secreted into the culture medium, with a yield of 25mg/L as shown by SDS-PAGE and Western blotting. The expressed hTumstatin was purified to more than 85% purity using a simple one-step SP-Sepharose cation exchange chromatography. The MTT and chick chorioallantoic membrane assay showed that the yeast produced hTumstatin could inhibit the proliferation of human umbilical vein endothelial cells and the neovascularization induced by bFGF. Hoechst 33258 fluorescent staining also demonstrated the apoptotic change in endothelial cellular nuclear morphology.
Subject(s)
Humans , Angiogenesis Inhibitors , Metabolism , Autoantigens , Genetics , Metabolism , Cell Proliferation , Cells, Cultured , Collagen Type IV , Genetics , Metabolism , DNA, Complementary , Genetics , Electroporation , Endothelial Cells , Cell Biology , Pichia , Genetics , Metabolism , Recombinant Proteins , Genetics , Metabolism , Umbilical Cord , Cell BiologyABSTRACT
<p><b>AIM</b>To determine peimine and peiminine in Fritillaria drug simultaneously by RP-HPLC-ELSD.</p><p><b>METHODS</b>HPLC was carried out with a Waters Alliance, Model 2690, equipped with XTerra RP18 column (150 mm x 3.9 mm ID, 5 microm) and evaporated light scattering detector. The mobile phase (acetonitrile-10 mmol.L(-1) NH4HCO3 adjusted to pH 10.10 by ammonia solution) was eluted in gradient mode.</p><p><b>RESULTS</b>The recoveries of peimine and peiminine were 98.96% (n = 4), with RSD 1.01% and 98.40% (n = 4), with RSD 2.63%, respectively.</p><p><b>CONCLUSION</b>The method is simple, sensitive and reliable. It can be used for quantitative determination of Fritillaria drug.</p>