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1.
Article in Chinese | WPRIM | ID: wpr-928007

ABSTRACT

Monoterpenes are widely used in cosmetics, food, medicine, agriculture and other fields. With the development of synthetic biology, it is considered as a potential way to create microbial cell factories to produce monoterpenes. Engineering Saccharomyces cerevisiae to produce monoterpenes has been a research hotspot in synthetic biology. In S. cerevisiae, the production of geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) is catalyzed by a bifunctional enzyme farnesyl pyrophosphate synthetase(encoded by ERG20 gene) which is inclined to synthesize FPP essential for yeast growth. Therefore, reasonable control of FPP synthesis is the basis for efficient monoterpene synthesis in yeast cell factories. In order to achieve dynamic control from GPP to FPP biosynthesis in S. cerevisiae, we obtained a novel chassis strain HP001-pERG1-ERG20 by replacing the ERG20 promoter of the chassis strain HP001 with the promoter of cyclosqualene cyclase(ERG1) gene. Further, we reconstructed the metabolic pathway by using GPP and neryl diphosphate(NPP), cis-GPP as substrates in HP001-pERG1-ERG20. The yield of GPP-derived linalool increased by 42.5% to 7.6 mg·L~(-1), and that of NPP-derived nerol increased by 1 436.4% to 8.3 mg·L~(-1). This study provides a basis for the production of monoterpenes by microbial fermentation.


Subject(s)
Fermentation , Geranyltranstransferase/genetics , Monoterpenes/metabolism , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism
2.
Article in Chinese | WPRIM | ID: wpr-927947

ABSTRACT

Ginsenoside Rh_2 is a rare active ingredient in precious Chinese medicinal materials such as Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Panacis Quinquefolii Radix. It has important pharmacological activities such as anti-cancer and improving human immunity. However, due to the extremely low content of ginsenoside Rh_2 in the source plants, the traditional way of obtaining it has limitations. This study intended to apply synthetic biological technology to develop a cell factory of Saccharomyces cerevisiae to produce Rh_2 by low-cost fermentation. First, we used the high protopanaxadiol(PPD)-yielding strain LPTA as the chassis strain, and inserted the Panax notoginseng enzyme gene Pn1-31, together with yeast UDP-glucose supply module genes[phosphoglucose mutase 1(PGM1), α-phosphoglucose mutase(PGM2), and uridine diphosphate glucose pyrophosphorylase(UGP1)], into the EGH1 locus of yeast chromosome. The engineered strain LPTA-RH2 produced 17.10 mg·g~(-1) ginsenoside Rh_2. This strain had low yield of Rh_2 while accumulated much precursor PPD, which severely restricted the application of this strain. In order to further improve the production of ginsenoside Rh_2, we strengthened the UDP glucose supply module and ginsenoside Rh_2 synthesis module by engineered strain LPTA-RH2-T. The shaking flask yield of ginsenoside Rh_2 was increased to 36.26 mg·g~(-1), which accounted for 3.63% of the dry weight of yeast cells. Compared with those of the original strain LPTA-RH2, the final production and the conversion efficiency of Rh_2 increased by 112.11% and 65.14%, respectively. This study provides an important basis for further obtaining the industrial-grade cell factory for the production of ginsenoside Rh_2.


Subject(s)
Fermentation , Ginsenosides , Humans , Panax/genetics , Panax notoginseng , Saccharomyces cerevisiae/genetics , Uridine Diphosphate Glucose
3.
Article in English | WPRIM | ID: wpr-772771

ABSTRACT

Konjac glucomannan (KGM) is a water-soluble polysaccharide obtained from the roots and tubers of konjac plants. Recently, a degraded product of KGM, depolymerized KGM (DKGM), has attracted attention because of its low viscosity, improved hydrophily, and favorable physiological functions. In this review, we describe the preparation of DKGM and its prebiotic effects. Other health benefits of DKGM, covering antioxidant and immune activity, are also discussed, as well as its safety. DKGM could be a candidate for use as a tool for the treatment of various diseases, including intestinal flora imbalance, and oxidative- and immune-related disorders.


Subject(s)
Amorphophallus , Chemistry , Animals , Antioxidants , Therapeutic Uses , Humans , Hydrophobic and Hydrophilic Interactions , Immunologic Factors , Therapeutic Uses , Mannans , Therapeutic Uses , Plants, Medicinal , Chemistry , Polymerization , Prebiotics , Safety , Viscosity
4.
Chinese Medical Journal ; (24): 2658-2665, 2018.
Article in English | WPRIM | ID: wpr-775037

ABSTRACT

Background@#Treatment with the dipeptidyl peptidase-4 inhibitors (DPP4i) and angiotensin receptor blockers (ARBs) in patients with type 2 diabetic nephropathy (DN) has not been well characterized. This study aimed to assess the renoprotection of this combined treatment in DN patients.@*Methods@#A total of 159 type 2 DN patients from 2013 to 2015 were enrolled retrospectively from a prospective DN cohort at the National Clinical Research Center of Kidney Diseases, Jinling Hospital (China). Fifty-seven patients received DPP4i and ARB treatment, and 102 patients were treated with ARBs alone. All patients were followed up for at least 12 months. Statistical analyses were performed using Stata version 12.0.@*Results@#There were no significant differences at baseline for age, sex, body mass index, duration of diabetes, fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and estimated glomerular filtration rate (eGFR) between the two groups. Antihypertensive and antidiabetic medication use was similar in each group except calcium channel antagonists (P = 0.032). No significant changes in FBG and HbA1c were observed in the two groups after treatment. The eGFR decreased slower in the DPP4i + ARB group than in the ARB group at 12 months (Δ12 months: -2.48 ± 13.86 vs. -6.81 ± 12.52 ml·min·1.73m, P = 0.044). In addition, proteinuria was decreased further in the DPP4i + ARB group than in the ARB group after 24 months of treatment (Δ24 months: -0.18 [-1.00, 0.17] vs. 0.32 [-0.35, 0.88], P = 0.031). There were 36 patients with an eGFR decrease of more than 30% over 24 months. After adjusting for FBG, HbA1c, and other risk factors, DPP4i + ARB treatment was still associated with a reduced incidence of an eGFR decrease of 20% or 30%.@*Conclusions@#The combined treatment of DPP4i and ARBs is superior to ARBs alone, as evidenced by the greater proteinuria reduction and lower eGFR decline. In addition, the renoprotection of DPP4i combined with ARBs was independent of glycemic control.


Subject(s)
Aged , Angiotensin Receptor Antagonists , Therapeutic Uses , Diabetic Nephropathies , Drug Therapy , Dipeptidyl-Peptidase IV Inhibitors , Therapeutic Uses , Female , Humans , Losartan , Therapeutic Uses , Male , Middle Aged , Prospective Studies , Retrospective Studies
5.
Article in Chinese | WPRIM | ID: wpr-699378

ABSTRACT

Objective:To explore influence of amlodipine besylate tablet combined metoprolol on serum inflammatory factor levels and cardiac function in aged patients with essential hypertension(EH)complicated chronic heart failure(CHF).Methods:A total of 98 aged EH+CHF patients treated in our hospital from Jan 2014 to Oct 2015 were se-lected.Patients were randomly and equally divided into metoprolol group(received metoprolol based on routine treatment)and combined treatment group(received amlodipine besylate tablet based on metoprolol group),both groups were treated for six months.Serum levels of interleukin(IL)-1,IL-6 and tumor necrosis factor(TNF)-α, left ventricular end-diastolic dimension(LVEDd),left ventricular end-systolic dimension(LVESd)and left ventric-ular ejection fraction(LVEF)before and six months after treatment,and therapeutic effect were measured and compared between two groups.Results:Compared with before treatment,after six-month treatment,there were significant reductions in serum levels of IL-1,IL-6,TNF-α,LVEDd and LVESd,and significant rise in LVEF in two groups,P=0.001 all;compared with metoprolol group after six-month treatment,there were significant reduc-tions in serum levels of IL-1[(16.21 ± 1.45)ng/L vs.(12.34 ± 1.32)ng/L],IL-6[(46.21 ± 5.32)ng/L vs. (36.42 ± 4.51)ng/L],TNF-α[(23.12 ± 2.14)ng/L vs.(19.25 ± 2.41)ng/L],LVEDd[(51.68 ± 4.74)mm vs. (42.31 ± 4.25)mm]and LVESd[(46.27 ± 4.78)mm vs.(39.06 ± 4.35)mm],and significant rise in LVEF [(43.26 ± 4.37)% vs.(52.95 ± 5.18)%]in combined treatment group,P=0.001 all.Total effective rate of com-bined treatment group was significantly higher than that of metoprolol group(93.88% vs.79.59%),P= 0.037. There was no significant difference in incidence rate of adverse reactions between two groups,P=0.505. Conclu-sion:Amlodipine besylate combined metoprolol possesses significant therapeutic effect on aged EH + CHF patients, which may reduce serum inflammatory factor levels,relieve ventricular remodeling and improve cardiac function.

6.
Article in Chinese | WPRIM | ID: wpr-698542

ABSTRACT

BACKGROUND: Our research team has confirmed that compared to the adenoviral vector, transfection by lentiviral vector to rabbit bone marrow mesenchymal stem cells (BMSCs) is more effective that the expression of enhanced green fluorescent protein (EGFP)/bone morphogenetic protein 2 (BMP-2) can be persistent for a longer term. But further investigations are needed to explore whether BMSCs transfected with hBMP-2 through lentivirus combined with demineralized bone matrix (DBM) can promote bone defect repair. OBJECTIVE: To evaluate the effect of lentivirus-mediated hBMP-2/BMSCs/DBM (LV-hBMP-2/BMSCs/DBM) on the repair of large-segmental femoral defects and to explore the new treatments for large-segmental femoral defects. METHODS: Large-segmental bone defect models were made in the right femur of 48 New Zealand white rabbits by cutting the middle femoral bone and steel plate fixation. Then, animal models were randomly divided into four groups (n=12 per group) and implanted with nothing (control), DBM, hBMP-2/DBM, and LV-hBMP-2/BMSCs/DBM. Three rabbits from each group were sacrificed at 2, 4, 8 and 12 weeks after surgery to evaluate the repairing effect of femoral defects through hematoxylin-eosin staining, biomechanical analysis and radiological examination. RESULTS AND CONCLUSION: X-ray results revealed that osteotylus formed in all the four groups to different extents, and Lane - Sandhu X-ray scores were ranked as follows: control group < DBM group < hBMP-2/DBM group < LV-hBMP-2/BMSCs/DBM group (P < 0.05). Findings from the three-point bending test showed that the maximum load of the LV-hBMP-2/BMSCs/DBM group was significantly higher than that of the hBMP-2/DBM group, but is still lower than that of the normal femur at 8 and 12 weeks after modeling (P < 0.05). Hematoxylin-eosin staining results revealed that a few trabecular bones arranged disorderedly and a large amount of fibrous tissues in the control group; the DBM scaffold was basically degraded in the DBM group presenting with partially disordered trabecular bones and a large amount of fibrous tissues; the trabecular bones in the bone defect area were basically connected into line to start the shaping of the cortical bone, and recanalization of the medullary cavity was insignificant in the hBMP-2/DBM group; new cortical bone formed in the bone defect area and the medullary cavity was recanalized in the LV-hBMP-2/BMSCs/DBM group. These findings indicate that LV-hBMP-2/BMSCs/DBM can produce a large amount of calluses, promote formation of new cortical bone, and promote bone conduction, bone induction and osteogenesis after implantation into the bone defect; and this material has good repairing effect on large-segmental femoral defects of rabbits.

7.
Progress in Modern Biomedicine ; (24): 4831-4836,4891, 2017.
Article in Chinese | WPRIM | ID: wpr-615164

ABSTRACT

Objective:The current study investigated the effects of nesfatin-1 in the hypothalamic paraventricular nucleus (PVN) on gastric motility and the regulation of the lateral hypothalamic area (LHA).Methods:The projection of nerve ?ber and expression of nesfatin-1 were observed by retrograde tracing and fluo-immunohistochemistry staining;The nuclei microinjection and nuclei electrical stimulation,extracellular discharges of single unit neuron were used to observe the effects of nesfatin-1 on the GD neurons;Gastric motility recording in vivo were used to monitor the effects of nesfatin-1 on the amplitude of constriction and frequency of gastric motility in conscious rats.Results:Nesfatin-1 inhibited the majority of the GD-E neurons(1.97± 0.12 Hz vs.1.15± 0.07 Hz) and excited GD-I neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) in the PVN,which were weakened by oxytocin receptor antagonist H4928 (GD-E:1.38± 0.08 Hz,P<0.05 vs.nesfatin-1;GD-I:2.49± 0.15 Hz,P<0.05 vs.nesfatin-1).Gastric motility experiments showed that administration ofnesfatin-1 in the PVN decreased gastric motility.Retrograde tracing and immunofluorescent staining showed that nucleobindin-2/nesfatin-1 and fluorogold double-labeled neurons were observed in the LHA.Electrical LHA stimulation excited the firing rate of GD-responsive neurons (GD-E:2.06± 0.12 Hz vs.4.23± 0.21 Hz,GD-I:1.61± 0.09 Hz vs.4.83± 0.25 Hz) in the PVN.Pre-administration of an antinucleobindin-2/nesfatin-1 antibody in the PVN strengthened gastric motility,decreased GD-E neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) and excited the discharging of the GD-I neurons(4.15± 0.18 Hz vs.4.83± 0.25) induced by electrical stimulation of the LHA.Conclusion:Nesfatin-1 in the PVN could serve as an inhibitory factor to inhibit gastric motility,which might be regulated by the LHA.

9.
Article in English | WPRIM | ID: wpr-34085

ABSTRACT

OBJECTIVES: To investigate the surgical outcomes of different uvulopalatopharyngoplasty (UPPP). METHODS: All subjects underwent overnight polysomnography and were evaluated using the Epworth sleepiness scale (ESS), the Quebec sleep questionnaire and the snoring scale at the baseline and 3 and 12 months following operation. The primary endpoint was the overall effective rate representing the sum of the surgical success rate and effective rate. RESULTS: The overall effective rate at 12 months post surgery was 55.6% for simple UPPP, 95.8% for UPPP+GA, and 92.3% for UPPP+TBA. The surgical success rate at 3 and 12 months postoperation for UPPP+GA or UPPP+TBA was significantly higher than simple UPPP (P<0.05). Marked improvement was observed in all patients in the snoring scale score and the ESS score 3 and 12 months following surgery compared to the baseline (P<0.05 in all). CONCLUSION: UPPP, UPPP+GA, and UPPP+TBA are all effective in improving the surgical outcome of obstructive sleep apnea hypopnea syndrome (OSAHS) patients with multilevel obstruction. UPPP+TBA appears to be the most effective in treating OSAHS patients.


Subject(s)
Humans , Polysomnography , Prospective Studies , Quebec , Sleep Apnea Syndromes , Sleep Apnea, Obstructive , Snoring , Tongue , Surveys and Questionnaires
10.
Cancer Research and Clinic ; (6): 760-762,770, 2015.
Article in Chinese | WPRIM | ID: wpr-603248

ABSTRACT

Objective To analyze the clinical characters,treatment effects and influential factors of gastrointestinal stromal tumor patients with hepatic metastases,and to explore the reasonable clinical treatment protocols.Methods The clinical data of 24 gastrointestinal stromal tumor patients with hepatic metastases who were admitted to Peoples Liberation Army Hospital from April 2007 to April 2010 were retrospectively analyzed.Results The 1-,2-,3-and 4-year cumulative survival rates of hepatic metastases patients with gastrointestinal stromal tumors were 91%,72 %,22 % and 6 %,respectively.The 1-,2-and 3-year survival rates of the patients treated with imatinib after operation were 91%,70 %,28 %,respectively,the median survival time was 26 months,the mortality rate was 27.3 % (3/11),and 9 cases got the tumor complete elimination.The 1-,2-and 3-year survival rates of the patients treated without imatinib after operation were 90 %,68 %,18 %,respectively,the median survival time was 24 months,the mortality rate was 14.3 % (2/13),and 12 cases got the tumor complete elimination.The recurrence rate after complete resection was 28.6 %(4/14),and the mortality rate was 7.1% (1/14).The mortality rate after incomplete resection was 40 % (4/10).Conclusion Surgical treatment of eliminating the primary and metastatic lesion,and adjuvant therapy of imtinib could prolong the survival time,reduce postoperative recurrence rate and improve the quality of life,which are the effective methods for the treatment of the gastrointestinal stromal tumor patients with hepatic metastases.

11.
Article in Chinese | WPRIM | ID: wpr-420463

ABSTRACT

Iatrogenic traumatic biliary stricture is one of the difficult points in the biliary surgery,and operation is the only definitive treatment. The operative opportunity,surgical procedure and techniques are important for the prognosis.From January 1998 to December 2011,173 patients with iatrogenic traumatic stricture were admitted to the Eastern Hepatobiliary Surgery Hospital. According to the Bismuth classification of traumatic biliary stricture,10 patients were in type Ⅰ,22 in type Ⅱ,87 in type Ⅲ,38 in type Ⅳ and 16 in type Ⅴ.Excision of the traumatic stricture with end-to-end anastomosis was performed on 19 patients.Of the 173 patients,154 were treated by Roux-en-Y duodenojejunostomy, and 8 of them received additional hemihepatectomy or partial hepatectomy.A total of 155patients were followed up,with a median time of 74 months,the total excellent and good rate was 94.8% ( 147/155 ).Surgery is the most effective therapy for iatrogenic traumatic biliary stricture. Optimal timing, reasonable surgical methods, strictly following the principle of biliary surgery and perfect operative skills are key points for a better prognosis.

12.
Chinese Medical Journal ; (24): 2795-2798, 2011.
Article in English | WPRIM | ID: wpr-292800

ABSTRACT

Large epiglottic cysts can block the glottis, leading to serious consequences. This condition presents a challenge in terms of airway management for anesthesiologists during induction of anesthesia. We report the use of a Shikani™ Seeing Optical Stylet combined with a Macintosh laryngoscope to aid tracheal intubation in seven patients with large epiglottic cysts. Use of this technique can avoid cyst rupture and allow smooth, safe intubation.


Subject(s)
Epiglottis , Pathology , Female , Fiber Optic Technology , Methods , Humans , Intubation, Intratracheal , Methods , Laryngeal Diseases , Pathology , Laryngoscopes , Male , Middle Aged
13.
Journal of Medical Biomechanics ; (6): E217-E224, 2011.
Article in Chinese | WPRIM | ID: wpr-804172

ABSTRACT

Objective To investigate effects of the cyclic stretch on rat bone marrow mesenchymal stem cells (rBMSCs) proliferation and the related signal molecules. Methods The cyclic mechanical stretching apparatus was used to study the effects of the cyclic stretch on the growth of rBMSCs with different parameters. The expression of c fos was measured by RT PCR at mRNA level and the levels of phosphorylated extracellular signal regulated kinase 1/2 (p-ERK1/2), total ERK1/2 (t-ERK1/2) and c-fos proteins were detected by Western blotting. Results The proliferation of rBMSCs could be stimulated by the cyclic stretch, and the number of cells was increased to 1.4 times compared with that of the control group at 1 Hz, 10% strain for 15 min and then being static for 6 h. Moreover, the expressions of c-fos mRNA and protein levels were increased significantly after the stimulation of the stretch. On the other hand, it was found that the cyclic stretch could remarkably enhance the phosphorylation of ERK1/2, but did not influence the level of t-ERK1/2. After treated with PD98059, the proliferation of rBMSCs stimulated by the cyclic stretch was inhibited prominently in a dose dependent manner and the expressions of p-ERK1/2, as well as c-fos at gene and protein levels were sharply down regulated. Conclusions ERK1/2 signal molecules play an important role in regulating the cyclic stretch on the proliferation of rBMSCs and transcription factors may take part in the regulations during this progress.

14.
Article in Chinese | WPRIM | ID: wpr-347533

ABSTRACT

<p><b>OBJECTIVE</b>To study the status of iron metabolism and erythropoietic proliferation in children with various genotypes of thalassemia.</p><p><b>METHODS</b>Serum concentrations of ferritin (SF), transferrin receptor (sTfR) and erythropoietin (EPO) were measured in 158 children with thalassemia. The differences in the concentrations of the three indices among children with different genotypes of thalassemia were compared. The correlations of the hemoglobin level with sereum SF, sTfR and EPO levels were assessed.</p><p><b>RESULTS</b>Among the 158 children with thalassemia, 52(32.9%) were diagnosed with alpha-thalassemia minor, 27(17.1%) with HbH disease, 59(37.4%) with beta-thalassemia minor, 13(8.2%) with beta-thalassemia major, and 7(4.4%) with combining alpha beta thalassemia. The SF levels in children with HbH disease or beta-thalassemia major were significantly higher than those in the other thalassemia groups (P<0.01). The sTfR levels in children with beta-thalassemia major were the highest when compared with those in the other thalassemia groups (P<0.05). The EPO levels in children with beta-thalassemia major were also the highest when compared with those in the other thalassemia groups (P<0.01). There was a negative correlation between hemoglobin and EPO levels in children with HbH disease (r=-0.656, P<0.01) and beta-thalassemia major (r=-0.641; P<0.05).</p><p><b>CONCLUSIONS</b>The status of iron metabolism and erythropoietic proliferation is different in children with different genotypes of thalassemia. A combined measurement of SF, sTfR and EPO may reflect the status of erythropoietic proliferation.</p>


Subject(s)
Adolescent , Child , Child, Preschool , Erythropoiesis , Erythropoietin , Blood , Female , Ferritins , Blood , Genotype , Humans , Infant , Iron , Metabolism , Male , Receptors, Transferrin , Blood , Thalassemia , Blood , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-597504

ABSTRACT

Objective To observe the biological activities of human doppel (Dpl) protein transiently expressed and Dpl-like protein PrPΔ32-121 on a human neuroblastoma cell line SH-SY5Y. Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrPΔ32-121 fragment were generated by PCR. The expression and location of Dpl and PrPΔ32-121 post-transfection were observed by IFA. The cytotoxicity was measured by MTT analysis. Cellular apoptosis was investigated by flow cytometry and Western blot. Results Both Dpl and PrPΔ32-121 protein were expressed and mainly located on the cell membrane. Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection. Meanwhile, more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrPΔ32-121 expressing plasmids. Conclusion Dpl protein transiently expressed and PrPΔ32-121 can lead to the similar neural cytotoxicity, probably triggering the cell apoptosis program.

16.
Chinese Journal of Virology ; (6): 368-375, 2009.
Article in Chinese | WPRIM | ID: wpr-297946

ABSTRACT

Recently, much work has been devoted to study MD-induced oncogenesis and the genes involved in this process. Among many genes in the MDV genome, several genes such as Meq, RLORF4, RLORF12, and 132bpr have been considered recently associated with virulence of MDV. In this paper, primers of Meq, RLORF4, RLORF12 and 132bpr genes were designed and synthesized, based on the published whole genome sequence of MDV strain GA. The genes of Meq, RLORF4 and RLORF12 from four Chinese epidemic MDV strains highly passaged on chicken embryo fibroblast (CEF), i. e. L-SYp85C, L-MSp75C, L-CZp75C, and L-ZYp75C, as well as their corresponding parent strains, i. e. L-SY, L-MS, L-CZ, and L-ZY, the reference virulent strain J-1 and the vaccine strain 814 were amplified by PCR respectively. Then the PCR products of interest were cloned and sequenced respectively. The results of sequence comparison and analysis of Meq genes in the study indicated that Meq genes from the two strains L-ZYp75C and L-CZp75C contained single nucleotide insertion and deletion. The Meq gene from strain L-ZYp75C contained an extra cytidine (C) insertion at nucleotide position 529 and a single thymidine (T) deletion at nucleotide position 602, resulting in a frameshift mutation. And this frameshift mutation could lead to changes in deduced amino acid sequence from position 177 to 200 of Meq gene. The extra C insertion at nucleotide position 625 in Meq gene of strain L-CZp75C was also predicted to cause frameshift mutation in three overlapping genes (Meq, RLORF6 and 23KD genes). The comparison of nucleotide sequences of RLORF4 genes in the study revealed that the RLORF4 gene of strain L-SYp85C contained a fragment deletion in Open Reading Frame (ORF) from nucleotide position 215 to 265, resulting in 17 amino acids deletions, which were not found in other sequenced strains. Comparison of nucleotide sequences of RLORF12 genes in the study revealed several mutations. The RLORF12 gene of strain L-MSp75C contained a single T deletion at nucleotide position 67 and of 814 vaccine strain a large fragment deletion from nucleotide position 18 to 86, both of the deletions located in Origin of replication site (Ori) of MDV genome. But strain L-ZYp75C possessed an unique "TGTTGGG" deletion in its RLORF12 gene. When the four Chinese epidemic MDV strains were serially passaged on CEF, the number of copies of the 132bp repeats increased from 2 to more than 10 copies. All of above results indicated that deletion and/or insertion mutation occurred in Meq, RLORF4, RLORF12 and 132bpr after serial passage of these four Chinese epidemic MDV strains on CEF.


Subject(s)
Amino Acid Sequence , Animals , Cells, Cultured , Chick Embryo , Chickens , DNA Mutational Analysis , Fibroblasts , Marek Disease , Genetics , Virology , Molecular Sequence Data , Mutation , Open Reading Frames , Genetics , Sequence Homology, Amino Acid , Viral Proteins , Chemistry , Genetics
17.
Article in Chinese | WPRIM | ID: wpr-304606

ABSTRACT

<p><b>OBJECTIVE</b>To identify the relationship between the expression of alpha and beta-isoforms of glucocorticoid receptors (GR) in peripheral blood mononuclear cells (PBMC) and glucocorticoid resistance in children with idiopathic thrombocytopenia purpura (ITP).</p><p><b>METHODS</b>Real-time PCR was used to detect the expression of GR alpha and GR beta mRNA in PBMC from 30 children with ITP (glucocorticoid-sensitive, n=18; glucocorticoid-resistant, n=12) and 10 healthy children (control group). Enzyme immunoassay was used to measure plasma levels of total glucocorticoids.</p><p><b>RESULTS</b>There were no significant differences in PBMC GR alpha mRNA levels among the glucocorticoid sensitive, the glucocorticoid-resistant and the control groups. Compared with the glucocorticoid-sensitive and the control groups, the expression of GR beta mRNA in the glucocorticoid-resistant group was significantly up-regulated (p<0.01). Plasma total glucocorticoids level in the glucocorticoid-resistant group was found to be much higher than that in the glucocorticoid-sensitive and the control groups (p<0.01).</p><p><b>CONCLUSIONS</b>The up-regulated expression of GR beta mRNA may associated with glucocorticoid resistance in children with ITP.</p>


Subject(s)
Child , Child, Preschool , Drug Resistance , Female , Glucocorticoids , Pharmacology , Humans , Male , Purpura, Thrombocytopenic, Idiopathic , Blood , Drug Therapy , RNA, Messenger , Receptors, Glucocorticoid , Blood , Genetics
18.
Article in Chinese | WPRIM | ID: wpr-329549

ABSTRACT

Objective To investigate the epidemiological,genealogic characteristic,familial history of the families with fatal familial insomnia,its clinical and pathological features as well as the heredity rule of related genes.Methods 135 familial members of 7 eras were studied.Vein blood samples from patients as well as from some familial members were collected.PRNP gene was studied with PCR,its serial was determined and then authenticated with Nsp I.Brain tissue was obtained for neuropathological test and PrPSc test with Western blot method.Results Clinical symptoms of the 2 diagnosed cases were typical.11 familial members died of similar neural disease.32 samples of their familial members,codon at D178N of PRNP of 11 members was mutated,with mutation rate as 34.38% while D129N showed as methionine.Brain tissue of both probands denaturalized into spongiform and the nerve fiber was absent but PrPSc protein was identified.Conclusion Genealogy was described in the family with fatal familial insomnia since the patients had typical clinical symptoms and pathological characteristics.It seemed necessary to confirm cases of fatal familial insomnia and their genealogy with epidemiological data and to investigate its gene characteristics as well as with neuropathological and Western blot tests.

19.
Chinese Journal of Virology ; (6): 202-207, 2009.
Article in Chinese | WPRIM | ID: wpr-334750

ABSTRACT

In human prion diseases, phosphorylated-tau deposition has been described in a rare genetic form, Gerstmann-Straussler-Scheinker disease, but is not considered part of the neuropathological picture of Creutzfeldt-Jakob disease. To investigate the possible changes of tau and phosphorylated tau (Ser396/Ser404) in transmissible spongiform encephalopathies (TSEs), the expressions and transcriptions of above biological factors in the brain tissues of 263K- and 139A-infected hamsters were evaluated by Western blots and Real Time PCR, respectively, followed by quantitative analyses of immunoblot images and relative transcriptional levels compared with normal animals. The contents of total tau increased, but phosphorylated tau at Ser396 and Ser404 decreased, regardless of the types of scrapie agents and clinical incubations. Transcriptions of two tau isoforms were also markedly increased. These findings suggested that dephosphorylation of tau at Ser396/Ser404 was a illness-correlative phenomenon in TSEs. Alterations of tau and phosphorylated tau (Ser396/Ser404) were either intermediate or consequent events in TSE pathogenesis and proposed the potential linkage of these bioactive proteins with the pathogenesis of prion diseases.


Subject(s)
Animals , Blotting, Western , Brain , Metabolism , Cricetinae , Gene Expression Regulation , Physiology , Phosphorylation , Polymerase Chain Reaction , PrPSc Proteins , Virulence , Prion Diseases , Metabolism , tau Proteins , Metabolism
20.
Article in Chinese | WPRIM | ID: wpr-317281

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and reliability of genioglossus advancement and hyoid suspension with non-trephine technic (GAHM) and uvulopalatopharyngoplasty (UPPP) for surgical obstructive sleep apnea-hypopnea syndrome treatment.</p><p><b>METHODS</b>Twenty-six patients (4 female and 22 male) were classified as moderate and severe cases in terms of apnea hypopnea index (AHI) and Friedman classification in the present study. All cases patients underwent genioglossus advancement and hyoid suspension with non-trephine technic Uvulopalatopharyngoplasty UPPP. Pre- and postoperative polysomnography and Epworth sleepiness scale were performed to assess the therapeutic outcomes.</p><p><b>RESULTS</b>The whole operation time ranged from 120-180 minutes. The average amount of bleeding in genioglossus advancement was about 50-100 ml. There were no severe complications during and after the operation. All the cases were followed up to one year. After surgery, AHI was decreased in both group cases, (42.9 +/- 6.6 vs 16.2 +/- 5.7) in the severe group, and 21.3 +/- 4.4 vs 11.3 +/- 5.2 (x(-) +/- s) in the moderate group. With success defined as AHI decreased by more than 50 per cent after surgery, the total success rate in moderate and severe group was 83.3% and 91.7%, respectively (P < 0.01). Furthermore, the success rate of the moderate group was higher than those of the severe group (P < 0.01).</p><p><b>CONCLUSION</b>GAHM combined with UPPP may be beneficial for the moderate and severe OSAHS patients with oropharyngeal and hypopharyngeal obstruction.</p>


Subject(s)
Adult , Chin , General Surgery , Female , Humans , Hyoid Bone , General Surgery , Male , Middle Aged , Otorhinolaryngologic Surgical Procedures , Methods , Palate, Soft , General Surgery , Pharynx , General Surgery , Sleep Apnea, Obstructive , General Surgery , Tongue , General Surgery , Uvula , General Surgery
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