Objective To set up a method of detecting platelet associated antibody(PA Ig) by flow cytometry(FCM),and to investigate the value of FCM method in the diagnosis of idiopathic thrombocytopenic purpura(ITP).Methods We detected the platelet associated antibody of 20 healthy donors and 10 patients with ITP.These samples were also measured with ELISA.The results of FCM were compared with the results of ELISA.Results By FCM,the measured value of PA IgG,PA IgA and PA IgM,from 20 normal dornors,were 2.56?2.08%、13.75?5.98% and 13.90?7.28%,respectively.In patients with ITP,PA IgG,PA IgA and PA IgM were 28.13?17.74%、25.07?16.84% and 23.22?17.85%.Conclusion FCM can be a rapid,sensitive and new method in detecting platelet autoantibody,and has better potential than ELISA.

Optically active form of alpha-cyano-3-phenoxybenzyl (CPB) alcohol, building block of pyrethroid insecticides, was synthesized as its acetate by the combination of anion-exchange resin (D301)-catalyzed transcyanation between m-phenoxybenzaldehyde (m-PBA) and acetone cyanohydrin (AC), and lipase (from Alcaligenes sp.)-catalyzed enantioselective transesterification of the resulting cyanohydrin with vinyl acetate. Through optimizing technological conditions, the catalyzing efficiency was improved considerably compared to methods previously reported. Concentrations of CPB acetate were determined by gas chromatograph. The enantio excess (e.e.) values of CPB acetate were measured by NMR (nuclear magnetic resonance) method. Effects of solvents and temperatures on this reaction were studied. Cyclohexane was shown to be the best solvent among the three tested solvents. 55 degrees C was the optimal temperature for higher degree of conversion. External diffusion limitation was excluded by raising the rotational speed to 220 r/min. However, internal diffusion could not be ignored, since the catalyst (lipase) was an immobilized enzyme and its particle dimension was not made small enough. The reaction rate was substantially accelerated when the reactant (m-PBA) concentration was as high as 249 mmol/L, but decreased when the initial concentration of m-PBA reached to 277 mmol/L. It was also found that the catalyzing capability of recovered lipase was high enough to use several batches. Study of the mole ratio of AC to m-PBA showed that 2:1 was the best choice. The strategy of adding base catalyst D301 was found to be an important factor in improving the degree of conversion of the reaction from 20% to 80%. The highest degree of conversion of the reaction has reached up to 80%.
Alcaligenes ; enzymology ; Benzaldehydes ; chemistry ; Combinatorial Chemistry Techniques ; methods ; Lipase ; chemistry ; Nitriles ; chemistry ; Organic Chemicals ; chemistry ; Phenyl Ethers ; chemical synthesis ; Technology, Pharmaceutical ; methods

OBJECTIVETo study the effect of Danshensu (DSS) and Ligustrazine (TMZ), the extracts of Chinese herbs for promoting blood circulation, on angiotensin II (Ang II) induced myocardial hypertrophy and its related genes, and to explore the mechanisms of inhibitory effect.

METHODSAdopting one-step method, the total RNA of myocardial cells was extracted by TRIzol reagent. Then the expression of ANP and beta-actin mRNA, as symbol of myocardial cells, were detected by RT-PCR.

RESULTSMolecular biological research showed that Ang II could significantly increase the expression of ANP mRNA in myocardial cells (P < 0.01), which could be significantly inhibited by Losartan (P < 0.01), both DSS and TMZ had the inhibitory effect (P < 0.05). Ang II could increase beta-actin mRNA expression in myocardial cells simultaneously, Losartan, DSS and TMZ could also significantly inhibit it (P < 0.05).

CONCLUSIONThe effective ingredients of Chinese herbs for promoting blood circulation, DSS and TMZ, have the effect of inhibiting the hyper-expression of ANP and beta-actin induced by Ang II, and preventing myocardial hypertrophy, therefore, it could be used to prevent and treat cardiomegaly.

Angiotensin II ; Animals ; Animals, Newborn ; Atrial Natriuretic Factor ; biosynthesis ; genetics ; Cardiomegaly ; chemically induced ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Lactates ; pharmacology ; Male ; Myocytes, Cardiac ; cytology ; metabolism ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar

OBJECTIVESTo elucidate the possible involvement of monoamine neurotransmitters in the development of neurobehavioral damage produced by acrylonitrile in drinking water in male rat brains.

METHODSTotally 30 male SD rats were randomly divided into three groups, the control group (n = 10), low dosage group (n = 10), and high dosage group (n = 10), which were respectively administered 0 mg/L, 50 mg/L, 200 mg/L acrylonitrile (AN) in drinking water. The treatment was lasted for 12 weeks. Seven animals were randomly selected from each group for determination of monoamine neurotransmitters in striatum and cerebellum by high performance liquid chromatography with electrochemical detector and activities of monoamine oxidase in cortex.

RESULTSThe contents of dopamine in the striatum of low and high dosage groups were decreased to (2.2 +/- 0.7) and (3.2 +/- 2.0) microg/g wet tissue, respectively, and compared with that of control group (9.0 +/- 4.2) microg/g wet tissue, the differences were statistically significant. There were no statistical differences among the contents of dopamine in the cerebellum of all rats, and the levels of 3,4-dihydroxyphenylacetic acid (DOPAC), the major metabolite of dopamine in the cerebellum were (186 +/- 41), (245 +/- 90) and (115 +/- 65) ng/g wet tissue in the control, low and high dosage groups, respectively and in low-dosage group they were significantly higher than those in other groups. There was dosage-dependently decreasing of the contents of serotonin of striatum in the control (249 +/- 34) ng/g wet tissue, low dosage (155 +/- 95) ng/g wet tissue and high dosage groups (128 +/- 101) ng/g wet tissue.

CONCLUSIONThis study underlines the importance of alterations in the monoamine neurotransmitters system as a possible causative mechanism behind the behavioural and functional changes produced by acrylonitrile.

Acrylonitrile ; administration & dosage ; toxicity ; Animals ; Biogenic Monoamines ; metabolism ; Brain ; drug effects ; metabolism ; Carcinogens ; administration & dosage ; toxicity ; Cerebellum ; drug effects ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Corpus Striatum ; drug effects ; metabolism ; Dopamine ; metabolism ; Dose-Response Relationship, Drug ; Drinking ; Male ; Neostriatum ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism

Objective:To describe the implementation status of the combination of medical-nursing and health care service and to analyze institutional dilemma,so as to put forward the corresponding policy recommendations.Methods:Based on the field investigation of 21 cities and the questionnaire survey of nursing homes,the implementation of support policy and the provision of the combination of medical-nursing and health care service were analyzed.Results:Relevant policy documents were issued,and the combination of medical care and nursing services were provided in the nursing home.However,the policy execution documents were more superficial and less practical.At the same time,although non-governmental forces was the main provider of the combination of medical care and nursing service.The medical care and nursing service were still in separation state.Conclusion:The mechanism and the reason of "difficult medical care and nursing services" were the existence of administrative isolated island between departments and the lack of incentive mechanism.

OBJECTIVETo study the relationship between ultrasonic bone mineral density (BMD) of calcaneus and age, height, and body weight, and to establish the normal reference value for stiffness index (SI) of healthy subjects in Guangxi autonomous region and provide scientific foundation for the diagnosis and prevention of osteoporosis.

METHODSSI of calcaneus in 1 816 healthy adults aged 11-90 years (775 men, 1 041 women) were measured with quantitative ultrasound. According to their ages, all the men and women were divided into 13 groups by 5-year age group.

RESULTSSI reached peak values in 21-25 year group in men (range: 111.45 +/- 21.19) and in 16-20 year group in women (range: 101.26 +/- 17.51). Then the SI value began to decline with aging. The SI featured by a typical dual-peak curve in women and the decrease rate of SI was faster in women than in men over 61 years. Multivariate linear regression analysis showed significantly negative correlation between SI and age (P < 0.001), positive correlation between SI and body weight (P < 0.05 for men; P < 0.01 for women), and positive correlation between SI and height in women (P < 0.01).

CONCLUSIONSI correlates with age, height, and weight. The acquired SI value may provide a reference for the diagnosis of osteoporosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Density ; Calcaneus ; diagnostic imaging ; Child ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Osteoporosis ; diagnosis ; epidemiology ; Ultrasonography

Objective To evaluate the clinical features and clinical outcome of liver, kidney and coagulation injury in patients with novel coronavirus pneumonia (COVID-19), especially in those elderly COVID-19 patients. Methods Clinical data of 72 COVID-19 patients admitted to the Shanghai Public Health Clinical Center from January 20, 2020 to April 20, 2020 was collected, including demographic data, clinical symptoms, laboratory testing results, underlying comorbidities, discharge and death, and then conducted the analysis stratified by clinical severity and age. Results Of the 72 COVID-19 patients, 27 were critically ill and 45 were mild.There were 31 elderly patients (≥60 years old) and 41 young patients (< 60 years old).Fever (83.3%), cough (36.1%) and fatigue (23.6%) were the most common clinical symptoms.The median age of the patients was 56.1 years (15-80 years); the mean age of mild and critically ill patients were 47.4 years and 64.6 years, respectively.The proportion of critically ill cases in the elderly group and the young group was 67.7% (21/31) and 14.6% (6/41), respectively.Organ injury in elderly patients included significantly increased aspartate aminotransferase, creatinine D-dimer, and FDP) (P < 0.05).Of the patients, the length of hospital stay of the elderly patients was significantly longer (P < 0.001), and the mortality increased significantly (P < 0.05). Conclusion Elderly COVID-19 patients have severe liver, kidney and coagulation injury, which may induce longer duration of hospital stay and increased mortality.Therefore, elderly COVID-19 patients may have worse clinical outcome.

OBJECTIVETo observe the dynamic time-phase expressions of key genes of brain-gut CaM signal pathway of spleen Qi deficiency rats and the intervention effect of Sijunzi decoction.

METHODMale Wistar rats were randomly divided into the normal control group, model 14 d, 21 d, 28 d groups, and Sijunzi decoction 14 d, 21 d, 28 d groups. Except for the normal control group, the remaining groups were included into the spleen Qi deficiency model with the bitter cold breaking Qi method (ig 7.5 g · kg⁻¹ · d⁻¹ of Rheum officinale, Fructus aurantii immaturus, Magnolia officinalis preparation) and the exhaustive swimming method. On the 7th day after the modeling, the Sijunzi decoction groups were orally administered with Sijunzi decoction 20 g · kg⁻¹ · d⁻¹. The expressions of key genes CaM/CaMK II of CaM signaling pathway in hippocampus and intestine at different time points by immunohistochemical method and Western blot. At the same time, the intervention effect of Sijunzi decoction on spleen Qi deficiency rats and its mechanism were analyzed.

RESULTSpleen Qi deficiency rats showed higher intestinal CaM/CaMK II expression and lower hippocampus CaM/CaMK II expression than normal rats (P < 0.05, P < 0.01). After the treatment of Sijunzi decoction, spleen Qi deficiency rats showed reduction in intestinal CaM/CaMK II expression and increase in hippocampus CaM/CaMK II expression (P < 0.05, P < 0.01).

CONCLUSIONThe formation of spleen Qi deficiency syndrome may be related to the high expression of CaM/CaMK II in small intestine tissues and its low expression in hippocampus tissues. Sijunzi decoction may achieve the therapeutic effect in spleen Qi deficiency syndrome by reducing the CaM/CaMK II expression in intestinal tissues and increasing it in hippocampus tissues.

Animals ; Brain ; drug effects ; enzymology ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Calmodulin ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Intestines ; drug effects ; enzymology ; metabolism ; Male ; Qi ; Rats ; Rats, Wistar ; Spleen ; drug effects ; Splenic Diseases ; drug therapy ; enzymology ; genetics ; metabolism

OBJECTIVETo investigate the influence of CD4+ CD25+ regulatory T cells(Treg cells) on mouse gastric cancer.

METHODSTreg cell in mouse spleen bearing gastric tumor was tested in different time points. Magic cell sorting(MACS) method was used to purify mouse Treg cells and the Treg cells were injected into mouse bearing gastric tumor with different dosage. After 3 weeks, the tumor size and tumor cell apoptosis rate were measured.

RESULTSTreg existed in normal mouse spleen with a rate of (3.86+/-0.07)%. In tumor model this percentage increased gradually and was (4.12+/-0.13)% after 3 weeks, which was significantly higher than that in control. When Treg cell applied in mouse reached 2.0 x 10(5), the tumor size enlarged significantly(P=0.013) and tumor cell apoptosis rate decreased significantly (P=0.012).

CONCLUSIONSTreg cell is associated with gastric cancer progress in mouse tumor model. Treg cell can promote gastric cancer growth and decrease tumor apoptosis. The anti- Treg GITR can improve anti- tumor effects.

Animals ; Apoptosis ; Female ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Spleen ; cytology ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance.

METHODSCD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines.

RESULTSThe expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji.

CONCLUSIONSmiR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.

B-Lymphocytes ; metabolism ; Cell Line, Tumor ; Cell Lineage ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; MicroRNAs ; metabolism