In industrial glutamate fermentation, intermitted feeding glucose with the help of off-line glucose measurement is generally necessary. This kind of feeding strategy could cause large variations in glucose concentration so that it is not favorable for the achievement of efficient and stable glutamate fermentation. Glutamate fermentation is characterized with typical non-growth association behavior, and during glutamate production phase glucose consumption is closely correlated with ammonia consumption. In this study, glucose concentration was controlled at various pre-determined levels by predicting glucose consumption amount and thus its concentration with the aid of on-line monitoring ammonia consumption. When glucose concentration was controlled around a lower level of 5 g/L~10 g/L, the final glutamate concentration could reach a relatively higher level of 80 g/L. In this way, the huge osmotic stress change due to the large glucose concentration variation with the intermitted feeding method could be avoided and the glutamate fermentation performance enhancement be expected.

ObjectiveTo investigate the effect of Jiuqiang Naoliqing(JNQ) on the TXA2 and PGI2 level in spontaneous hypertension rat (SHR) plasma.MethodsThe plasma was separated after the SHR and Wistar rats were treated with JNQ at the dose of 0.133g/kg,0.265g/kg,0.530g/kg and 1% carboxymethyl cellulose respectively for 5 weeks. The level of TXB2 and 6 keto PGF1α ,stable metabolin of TXA2 and PGI 2,in SHR plasma was tested by radioimmunoassay.ResultsThe level of TXB2 and the ratio of TXB2/6 keto PGF1α (T/P) in SHR plasma increased significantly(P<0.01),and there was no significant difference in the concentration of 6 keto PGF1α between Wistar rats and SHR plasma(P>0.05). JNQ could increase the generation of 6 keto PGF1α and decrease the level of TXB2 and T/P in SHR plasma after treated with different dosages for 5 weeks.ConclusionJNQ may improve the balance between TXA2 and PGI2 in SHR plasma.

Glutamate dehydrogenase (GDH) is a key enzyme in the biosynthesis of glutamate. The GDHs from Corynebacterium glutamicum S9114 the most commonly used strain in glutamate fermentation, were purified and their molecular structures and properties characterized. The coenzymes were also studied in the hope to increase glutamate production. Cells were harvested at mid-exponential phase by centrifugation and washed with Tris-HCl buffer containing DTT and EDTA (pH 7.5). The cells were then disrupted using a French pressure cell press and the supernatant was collected by centrifugation. The extract was concentrated by 70-fold using the AKTA-100 FPLC system employing a DEAE-cellulose ion exchange column, a hydrophobic interaction chromatography (HIC) and Sephadex G-200 gel filtration. The purified extracts contained NADPH-dependent GDH and NADH-dependent GDH. Both of the enzymes were highly specific for the coenzymes. The molecular masses of the NADPH-dependent GDH and its subunit were 188kD and 32kD respectively, suggesting the enzyme is a homo-hexamer. Our data reported for the first time the presence of NADH- dependent GDH in Corynebacterium glutamicum S9114, similar to other microorganisms containing both GDHs. The NADPH-dependent and NADH-dependent GDH in Corynebacterium glutamicum S9114 may participate in the assimilation and dissimilation of ammonia respectively. The absorptions of NADPH-dependent GDH was very weak at 280nm but very high at 215nm, suggesting a low phenylalanine and tyrosine content in the enzyme.
Chromatography, Gel ; Chromatography, Ion Exchange ; Corynebacterium glutamicum ; enzymology ; Glutamate Dehydrogenase ; isolation & purification ; metabolism ; Molecular Weight ; NADP ; metabolism ; Substrate Specificity

OBJECTIVETo evaluate the effect of JAK/STAT signaling pathway activation on the transdifferentiation and secretion of transforming growth factor-beta1 (TGF-beta1) induced by high glucose in renal proximal tubular epithelial cells.

METHODSHuman kidney cells (HKC) were cultured and then divided into four groups: low glucose (LG) group, high glucose (HG) group, high mannitol (LG + M) group, and HG + AG490 group. Immunoprecipitation and Western blot analysis were used to determine the expression of tryosine phosphorylated Janus kinase 2 ( p-JAK2). The protein expressions of STAT1, STAT3, p-STAT1, and p-STAT3 and the expressions of alpha-SMA and E-Cadherin were observed by Western blot. The contents of TGF-B1, fibronectin and type I collagen in the supernatants of the cultured HKC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-beta1 mRNA was measured by reverse transcription and polymerase chain reaction (RT-PCR).

RESULTSCompared with LG group, the expressions of JAK2, p-STAT1, p-STAT3, and TGF-beta1, mRNA were significantly increased in HG group from 6 to 72 hours. Meanwhile, the contents of TGF-beta1 and collagen I in the supernatants and the expression of alpha-SMA increased and the expression of E-Cadherin decreased. The expressions of JAK2, p-STAT1, p-STAT3, and TGF-beta mRNA as well as the levels of TGF-beta1 and collagen I in the supernatant s in HG + AG490 group were significantly lower than in the HG group. The expressions of alpha-SMA and E-Cadherin were also decreased in HG + AG490 group.

CONCLUSIONActivation of JAK/STAT signaling pathway may be involved in the high glucose-induced transdifferentiation and overproduction of TGF-beta1, and ECM proteins in HKCs.

Cell Line ; Cell Transdifferentiation ; Epithelial Cells ; cytology ; metabolism ; Glucose ; metabolism ; pharmacology ; Humans ; Janus Kinases ; physiology ; Kidney Tubules, Proximal ; cytology ; metabolism ; STAT Transcription Factors ; physiology ; Signal Transduction ; Transforming Growth Factor beta1 ; biosynthesis ; secretion ; Urothelium ; cytology ; metabolism

BACKGROUNDHypotension induced by combined spinal epidural anesthesia in parturient with hypertensive disorders of pregnancy (HDP) can easily compromise blood supply to vital organs including uteroplacental perfusion and result in fetal distress. The aim of this study was to investigate whether the goal-directed fluid therapy (GDFT) with LiDCO rapid system can improve well-being of both HDP parturient and their babies.

METHODSFifty-two stable HDP parturient scheduled for elective cesarean delivery were recruited. After loading with 10 ml/kg lactated Ringer's solution (LR), parturient were randomized to the GDFT and control group. In the GDFT group, individualized fluid therapy was guided by increase in stroke volume (ΔSV) provided via LiDCO rapid system. The control group received the routine fluid therapy. The primary endpoints included maternal hypotension and the doses of vasopressors administered prior to fetal delivery. The secondary endpoints included umbilical blood gas abnormalities and neonatal adverse events.

RESULTSThe severity of HDP was similar between two groups. The total LR infusion (P < 0.01) and urine output (P < 0.05) were higher in the GDFT group than in the control group. Following twice fluid challenge tests, the systolic blood pressure, mean blood pressure, cardiac output and SV in the GDFT group were significantly higher, and the heart rate was lower than in the control group. The incidence of maternal hypotension and doses of phenylephrine used prior to fetal delivery were significantly higher in the control group than in the GDFT group (P < 0.01). There were no differences in the Apgar scores between two groups. In the control group, the mean values of pH in umbilical artery/vein were remarkably decreased (P < 0.05), and the incidences of neonatal hypercapnia and hypoxemia were statistically increased (P < 0.05) than in the GDFT group.

CONCLUSIONSDynamic responsiveness guided fluid therapy with the LiDCO rapid system may provide potential benefits to stable HDP parturient and their babies.

Adult ; Anesthesia, Epidural ; methods ; Anesthesia, Spinal ; methods ; Blood Pressure ; Cesarean Section ; methods ; Female ; Fluid Therapy ; methods ; Humans ; Hypertension, Pregnancy-Induced ; Infant, Newborn ; Isotonic Solutions ; Pregnancy ; Pregnancy Outcome

Objective:To analyze the trend relevant factors leading to death and their patterns over a 10-year period in inpatients with connective tissue diseases (CTDs).Methods:All clinical data about death in inpatients with CTDs were retrospectively reviewed between 2005 and 2014 at the Department of Rheumatology and Immunology in Xiangya Hospital of Central South University.Results:In the 10-year time period,the overall hospital mortality was 15.689‰.The disease itself accounted for 44.71% of the total causes of death,infection accounted for 42.94%,and comorbidities accounted for 12.35%.The constituent ratio of deaths and the average hospital mortality caused by the disease itself declined gradually year by year,and the constituent ratio of deaths caused by infection and comorbidities increased gradually year by year (P<0.05).In 2013-2014,infection was the leading cause of death,which accounted for 51.06%.The survival time for CTDs inpatients with interstitial lung disease (ILD) was shorter than that of CTDs inpatients without ILD,and even the risk of death was 1.722 times of the latter.The proportion of deaths caused by the disease itself was the highest in systemic sclerosis and systemic lupus erythematosus,that by infection was the highest in idiopathic inflammatory myopathy (IIM),and that by comorbidities was the highest in rheumatoid arthritis.Conclusion:The proportion of deaths and the hospital mortality in CTDs inpatients caused by the disease itself show a declining trend,while the proportion of deaths caused by infection and comorbidities increase.CTDs patients with ILD have shorter survival time and an increase in risk of death.

OBJECTIVETo study the relationship between blood pressure control status and patients' knowledge on hypertension prevention and control among hypertensive patients.

METHODSA total of 726 hypertensives were selected from four community health service centers (2 urban and 2 rural) in Beijing. All subjects were investigated by questionnaires and their blood pressures were measured at the same time.

RESULTSThe rate for blood pressure under control (< 140/90 mm Hg, 1 mm Hg = 0.133 kPa) in the rural and urban patients were 46.4% and 23.9% respectively. The control rate increased with the increase of patients' knowledge on prevention and control of hypertension in both urban and rural patients. The cumulative effect of knowledge on hypertension control status could contribute 30.0% to the difference in hypertension control rate between rural and urban patients.

CONCLUSIONPatients' knowledge on hypertension control was significantly related to the rate on hypertension control. Health education should be helpful to improve the rate on hypertension control.

Aged ; Blood Pressure ; physiology ; Blood Pressure Monitoring, Ambulatory ; China ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Hypertension ; physiopathology ; prevention & control ; therapy ; Logistic Models ; Male ; Middle Aged ; Outpatients ; education ; Patient Education as Topic ; organization & administration ; Rural Population ; Social Class ; Surveys and Questionnaires ; Treatment Outcome ; Urban Population

【Objective】 To study the mutation characteristics of Tyrosine hydroxyls（TH） gene in a pedigree with dopa-responsive dystonia（DRD）. 【Methods】 Extraction of genomic DNA from peripheral blood of a proband and his parents and two sisters using high- throughput sequencing （NGS） method were detected on 256 known pathogenicity genes associated with dystonia and dyskinesia.【Results】Mutations on tyrosine hydroxylase（TH）gene in the exon 14 and exon 9 were detected in the proband and his eldest sister in this pedigree. They had a complex heterozygosity of c.1481C > T（p.Thr494Met）and c.943G >A（p.Gly315Ser），and one heterozygous mutation was carried by parents respectively. The mutation was not detected in his second sister and 50 people with normal phenotype controls. 【Conclusion】 The mutations of TH gene c. 1481C > T（p.Thr494Met）and c. 943G > A（p.Gly315Ser）led to the gene abnormality in DRD family，and a new mutation of TH gene was found，which expanded the relationship between DRD genotype and clinical phenotype. It is vital that early accurate diagnosis and treatment of DRD is the key to improve prognosis.

This study is to establish a simple and practical co-culture method of cortical neurons and astrocytes of rats. The cortex of the new-born SD rats was digested by 0.125% pancreatic enzyme, and the differential adherence was applied to obtain the mixed cell suspension of neurons and astrocytes. A low concentration of cytarabine was used to inhibit the astrocytes in a moderate way to get neuronal and astrocyte co-culture. The morphological characteristics of the cells in different times were observed under the inverted microscope. The cells began to adhere the wall 2 h after the inoculation. Neurons and astrocytes grew in a good condition under the inverted microscope 9 days after the inoculation. The results of the immunofluorescence staining and Rosenfeld's staining indicated that the co-culture of neurons and astrocytes was successful and the ratio of neurons and astrocytes was close to 1:1. A new neurons and astrocytes co-culture method, which is simple and convenient, was successfully established. It will be an efficient method for the related researches about neuronal and astrocyte co-culture in vitro.
Animals ; Animals, Newborn ; Astrocytes ; cytology ; Cells, Cultured ; Cerebral Cortex ; cytology ; Coculture Techniques ; methods ; Female ; Male ; Neurons ; cytology ; Primary Cell Culture ; methods ; Rats ; Rats, Sprague-Dawley