Based on reservoir condition and fluid characteristics, the oil-degrading bacterial strain NX-2 was screened from Ma-2 fault block of Huabei oil field. Bacterial metabolism and the capability of improving oil property were evaluated on oxygen-deficient condition. At 87℃ which reservoir temperature is, artificial homogeneous core displacement experiment indicated the enhanced oil recovery of microbe was 7.1% higher than that of waterflooding. In experiment on individual well Ma-410, additional oil production of 669 tons was gained, and decreased water production of this trial well reached more than 3000 tons. These results demonstrated NX-2 could adapt to stratum conditions, enhance oil recovery and improve oil property as well.

Objective To evaluate the relationship between Alzheimer′s disease(AD)and its partial vascular risk factors,and investigate the effect of tanshinone ⅡA(Tan ⅡA)on experimental models of AD combined with chronic cerebral ischemia and its potential mechanism.Methods Eight hundred and forty-nine patients with dementia(549 having diagnosis of AD and 300 without AD),admitted to our hospital from 1975 to 2009,were collected in our study; univariate analysis was performed on the relation between AD and vascular risk factors.Besides that,Tg+ mice were employed in our study and randomly divided into 5 groups,namely,sham-operated group,vehicle group,low Tan ⅡA treatment group,medium Tan ⅡA treatment group and high Tan ⅡA treatment group; or these mice were randomly divided into sham-operated group,vehicle group,Tan ⅡA treatment group(treated with 10 mg/kg daily).Mouse models of AD and chronic cerebral ischemia were established,and Tan ⅡA treatment was given to the Tan ⅡA treatment group.The relationship between AD and vascular factors was assessed by means of analyzing the clinicopathological data of AD cohort.The changes of learning and memory abilities in the mouse models were detected by Morris water maze test.Enzyme-linked immuno sorbent assay(ELISA)was employed to detect the level of VEGF; the protein expressions of betaA4-amyloid precursor protein (APP),VEGF and VEGFR-1 were determined by Western blotting,and the mRNA expressions of APP,VEGF and VEGFR-1 were observed by quantitative RT PCR.The effect of Tan ⅡA on canaliculization of human umbilical vein endothelial cells(HUVECs)was also investigated fiom cellular level.Results AD was significantly positively correlated with such vascular risk factors as hypertension,diabetes mellitus,coronary disease,cerebrovascular disease and hyperlipemia(P＜0.05),while no correlation was noted between AD and pneumonia.The mice of the medium Tan ⅡA treatment group and high Tan ⅡA treatment group had obviously shortened times of searching the platform and swimming distance,prolonged latency of avoiding darkness,decreased frequency of wrong behaviors,and decreased level of VEGF as compared with the vehicle group(P＜0.05).The life span in mice of the Tan ⅡA treatment group (treated with 10 mg/kg daily)was prolonged for approximately 24 d as compared with that in the vehicle group; the expressions of APP and VEGF were down-regulated and that of VEGFR-1 in mice of the Tan ⅡA treatment group(treated with 10 mg/kg daily)was up-regulated as compared with those in the vehicle group(P＜0.05).The canaliculization of HUVECs was enhanced after incubation with Tan ⅡA for 48 h,followed by increase of VEGFR-1 expression.Conclusion AD is significantly correlated with its vascular factors.Tan ⅡA could improve the learning and memory abilities of dementia mouse through up-regnlation of VEGFR-1 expression and promotion of vascular integrity,indicating the crucial role of vascular factors in treatment of AD.

BACKGROUNDHuman amniotic epithelial cells (HAECs) are able to secrete biologically active neurotrophins such as brain-derived neurotrophic factor and neurotrophin-3, both of which exhibit trophic activities on dopamine neurons. Previous study showed that when human amniotic epithelial cells were transplanted into the striatum of 6-hydroxydopamine (6-OHDA)-induced Parkinson disease rats, the cells could survive and exert functional effects. The purpose of this study was to investigate the survival and the differentiation of human amniotic epithelial cells after being transplanted into the lateral ventricle of Parkinson's disease (PD) rats, and to investigate the effects of grafts on healing PD in models.

METHODSThe Parkinson's model was made with stereotactic microinjection of 6-hydroxydopamine (6-OHDA) into the striatum of a rat. The PD models were divided into two groups: the HAECs group and the normal saline (NS) group. Some untreated rats were taken as the control. The rotational asymmetry induced by apomorphine of the HAECs group and the NS group were measured post cell transplantation. The expression of nestin and vimentin in grafts were determined by immunohistology. Ten weeks after transplantation the density of tyrosine hydroxylase positive cells in the substantia nigra of the HAECs group, NS group and the untreated group was determined. The differentiation of grafts was determined by TH immunohistology. High performance liquid chromatography (HPLC) was used to determine monoamine neurotransmitter levels in the striatum.

RESULTSThe rotational asymmetry induced by apomorphine of the HAECs group was ameliorated significantly compared to the NS group two weeks after transplantation (P < 0.01). The grafts expressed nestin and vimentin five weeks after transplantation. TH immunohistochemistry indicated that the TH positive cells in the substantia nigra of the HAECs group increased significantly compared to the NS group (P < 0.01). Tyrosine hydroxylase (TH) positive cells in the substantia nigra of the HAEC group and the NS group were decreased compared to the untreated group (P < 0.01). Dopamine and DOPAC levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.05). Homovanillic acid (HVA) levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.01). In addition dopamine, DOPAC, and HVA levels in the striatum and dopamine levels in the cerebrospinal fluid of the HAECs group and the NS group were decreased compared to the untreated group (P < 0.05).

CONCLUSIONSHuman amniotic epithelial cells could be used to ameliorate the rotational asymmetry induced by apomorphine of the PD models. This could have been due to the increased content of dopamine and its metabolic products, DOPAC and HVA, in the striatum in the PD models.

Amnion ; cytology ; Animals ; Apomorphine ; pharmacology ; Chromatography, High Pressure Liquid ; Epithelial Cells ; cytology ; drug effects ; transplantation ; Female ; Homovanillic Acid ; metabolism ; Humans ; Immunohistochemistry ; Oxidopamine ; toxicity ; Parkinsonian Disorders ; chemically induced ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley

Aged ; Aged, 80 and over ; Alzheimer Disease ; blood ; immunology ; B-Lymphocytes ; immunology ; B7-1 Antigen ; blood ; CD28 Antigens ; blood ; Female ; Flow Cytometry ; HLA-DR Antigens ; blood ; Humans ; Killer Cells, Natural ; immunology ; Lymphocyte Subsets ; immunology ; Male ; T-Lymphocyte Subsets ; immunology

OBJECTIVESTo study the effects of fenvalerate exposure on the semen quality of occupational workers in order to find out the early effective biomarkers.

METHODSThirty-two male workers who exposed to fenvalerate and 46 male administrators in the office in a pesticide factory were selected as the exposure group and internal control group, respectively, and 22 male administrators in a center for disease control were served as the external control group. In order to evaluate the exposed levels, the concentration of fenvalerate, toluene and xylene in the ambient air of working place in these three groups were monitored simultaneously for three consecutive days. After the semen were collected according to the standard method, the workers' semen qualities were analysed with University of California at Davis (UCDavis) method and the sperm motility were evaluated using computer assisted sperm analysis(CASA).

RESULTSIn the exposure group, the concentrations of fenvalerate were significantly higher than those in the internal and external control group (P < 0.01), while no significant difference of the concentration on toluene or xylene was found (P > 0.05). Sperm motion parameters in the exposure group such as linearity(LIN), straightness(STR), and the sperm count were decreased significantly, and the abnormality rate of viscidity, coagulation and sperm count were increased significantly as compared with the internal and the external control groups(P < 0.05). Furthermore, the sperm progression and beat cross frequency (BCF) in the exposure group were also lower significantly than those in the external control group(P < 0.05), while the abnormality rate of sperm progression was increased significantly.

CONCLUSIONSIn such a low concentration, occupational exposure to fenvalerate can affect workers' semen quality, especially the sperm count and sperm movement ability.

Adult ; Humans ; Insecticides ; toxicity ; Male ; Nitriles ; Occupational Exposure ; adverse effects ; Pyrethrins ; toxicity ; Semen ; drug effects ; Sperm Count ; Sperm Motility ; drug effects

BACKGROUNDHuman amniotic epithelial cells (HAECs), which have characteristics of both embryonic and pluripotent stem cells, are therefore a candidate in cell therapy without creating legal or ethical problems. In the present study, we aimed to investigate the effects of intracerebroventricular transplantation of HAECs on doubly transgenic mice of Alzheimer's disease (AD) coexpressing presenilin-1 (PS1) and mutant Sweden amyloid precursor protein (APPswe) genes.

METHODSThe offspring mice genotypes were detected using PCR identification of APPswe and PS1 gene. The doubly transgenic (TG) mice (n = 20) and wild-type (WT) mice (n = 20) were randomly divided into two groups respectively: the transplantation group treated with HAECs and the control group with phosphate buffered saline. Six radial arm water maze test was used to assess the spatial memory in the TG and WT mice. Amyloid plaques and neurofibrillary tangles were analyzed using congo red and acid-silver methenamine staining respectively. Immunofluorescence cytochemistry was used to track the survival of HAECs. Immunohistochemistry was used to determine the expression of octamer-binding protein 4 (Oct-4) and Nanog in the HAECs. High performance liquid chromatography was used to measure acetylcholine in hippocampus. The density of cholinergic neurons in basal forebrain and nerve fibers in hippocampus was measured using acetylcholinesterase staining.

RESULTSAmyloid deposition occurred in hippocampus and frontal cortex in the double TG mice aged 8 months, but not in WT mice. The results also showed that transplanted HAECs can survive for at least 8 weeks and migrate to the third ventricle without immune rejection. The graft HAECs can also express the specific marker Oct-4 and Nanog of stem cell. Compared with the control group, transplantation of HAECs can not only significantly improve the spatial memory of the TG mice, but also increase acetylcholine concentration and the number of hippocampal cholinergic neurites.

CONCLUSIONSThese results demonstrate that intracerebroventricular transplantation of HAECs can improve the spatial memory of the double TG mice. The higher content of acetylcholine in hippocampus released by more survived cholinergic neurites is one of the causes of this improvement.

Acetylcholine ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; therapy ; Amnion ; cytology ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Epithelial Cells ; cytology ; transplantation ; Genotype ; Hippocampus ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Memory Disorders ; genetics ; metabolism ; therapy ; Mice ; Mice, Transgenic ; Nanog Homeobox Protein ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Polymerase Chain Reaction ; Presenilin-1 ; genetics ; metabolism

ObjectiveTo establish the determination for index components in benchmark samples of Erdongtang， and clarify the content and transfer rate rages of index components in 15 batches of benchmark samples， and to explore the quantity transfer of index components of decoction pieces to benchmark samples. MethodFifteen batches of benchmark samples were prepared， the contents of mangiferin， baicalin and glycyrrhizic acid were determined by high performance liquid chromatography （HPLC）-diode array detector （DAD）， the mobile phase was acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 10%-17%A； 10-25 min， 17%-19%A； 25-28 min， 19%-25%A； 28-45 min， 25%-33%A； 45-46 min， 33%-45%A； 46-60 min， 45%-55%A）， detection wavelength was set at 254 nm. Contents of timosaponin BⅡ and the sum of protoneodioscin and protodioscin were determined by HPLC-evaporative light scattering detector （ELSD）， the mobile phase was acetonitrile （A）-water （B） for gradient elution （0-20 min， 24%A； 20-25 min， 24%-27%A； 25-33 min， 27%-28%A； 33-36 min， 28%-90%A； 36-41 min， 90%-24%A）. ResultThe methodological verification of the established method was good， which could be used for determination of five index components in benchmark samples. The content ranges of mangiferin， baicalin， glycyrrhizic acid， timosaponin BⅡ， and the sum of protoneodioscin and protodioscin in 15 batches of benchmark samples of Erdongtang were 0.14%-0.23%， 2.40%-3.37%， 0.07%-0.44%， 0.43%-0.95%， and 0.15%-0.47%， the transfer rate ranges of them were 33.90%-52.15%， 84.46%-105.61%， 22.59%-93.86%， 38.07%-61.43%， and 53.28%-96.11%， respectively. ConclusionThe consistencies of transfer rate of mangiferin， baicalin， timosaponin BⅡ and the sum of protoneodioscin and protodioscin （except glycyrrhizic acid） between decoction pieces and benchmark samples of Erdongtang are good， indicates that the transfer rates of 4 index components are stable during the preparation process of benchmark samples， which can provide data support for research and development of the compound preparation of this formula.

The quality evaluation method for standard decoction of Chinese herbal slices is the basis for the quality evaluation of granules and preparations of classical formula(decoction)of traditional Chinese medicine. This study aimed to establish a method for the determination of quercetin-3-O-glucuronic acid in Nelumbinis Folium(NF)and its standard decoction, so as to provide reference for the quality control of NF and its standard decoction. Fifteen batches of representative NF were collected to prepare standard decoction, and the parameters of dry extract rate, transfer rate of index component, and pH value were calculated. HPLC was used to establish the content determination method for quercetin-3-O-glucuronic acid in NF and its standard decoction. The concentration range of quercetin-3-O-glucuronic acid in the standard decoction of NF was 1.09-3.06 g·L~(-1), while the concentration range of nuciferine was 0.01-0.17 g·L~(-1). The average extraction rate of NF standard decoction was(14.4±2.6)%, the average transfer rate of quercetin-3-O-glucuronic acid was(70.7±18.6)%, and the average transfer rate of nuciferine was(9.6±5.4)%. Compared with Nuciferine, quercetin-3-O-glucuronic acid had a high content and stable transfer rate in standard decoction, and was recommended to be the quality control marker for NF and its standard decoction. This paper establishes a quality evaluation method for NF standard decoction, and can provide reference for the quality control of all preparations derived from NF and its decoction.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Flowers/chemistry* ; Medicine, Chinese Traditional ; Nelumbo/chemistry* ; Quality Control

Objective:There were 92 kinds of compound preparations containing Ophiopogonis Radix in the 2015 edition of Chinese Pharmacopoeia， but there was no effective method to identify these compound preparations. Because Ophiopogonis Radix and Liriopes Radix are similar in appearance， it is easy to be confused in application. The aim of this study was to set up a thin layer chromatography （TLC） to identify compound preparations containing Ophiopogonis Radix and distinguish Ophiopogonis Radix and Liriopes Radix in the forms of decoction pieces and standard decoction. Method:In this study， decoction pieces of Ophiopogonis Radix and Liriopes Radix were collected and separately prepared as standard decoction. TLC was used to qualitatively identify decoction pieces and standard decoction of Ophiopogonis Radix and Liriopes Radix， and compound preparations containing Ophiopogonis Radix. In the TLC， the lower solution of chloroform-methanol-water （65∶35∶10） was selected as the developing agent and 10% sulfuric acid ethanol solution as the chromogenic agent. Result:The resolution of this TLC was good. Decoction pieces， standard decoction and preparations of Ophiopogonis Radix had the same characteristic strips， which were two bright white fluorescent strips under ultraviolet lamp （365 nm）. But these two characteristic strips were not existed in the TLC of decoction pieces and standard decoction of Liriopes Radix. The corresponding components of both of these two strips were identified as mixture containing saponins by LC-MSⁿ， including ophiopogonin Ra， Tb， ophiopogonin D'， borneol glycoside， ophiopogonin C and Liriope muscari baily saponins C. Conclusion:The established TLC method， which has significant advantages such as high specificity and sensitivity， can be applied to the characteristic identification of decoction pieces and standard decoction of Ophiopogonis Radix， the identification of compound preparations containing Ophiopogonis Radix， and the distinction of Ophiopogonis Radix and Liriopes Radix， thus serving as an effective method to qualitatively identify Ophiopogonis Radix and its compound preparations.