Objective To investigate the effects of isoflurane preconditioning on renal ischemia reperfusion (I/R) injury in rats and the role of TNF-α plays in the mechanism.Methods Male SD rats were used in the study.The animals were randomly divided into 3 groups ( n =12 each):shame operation group; I/R group; Isoflurane preconditioning group (inhaled 1.5％ isoflurane (1 MAC) for 30 min followed by 10 min washout before I/R).At 2 h reperfusion,blood samples were obtained for urea nitrogen (BUN) concentration and creatinine (Cr) content.The level of TNF-α in renal tissues were determined by enzyme-linked immunosorbent assay (ELISA).Observe the pathological changes in H.E.staining slides under microscope.Results BUN concentration and Cr content and the level of TNF-α in I/R group and isoflurane preconditioning group were significantly higher than in shame operation group[ BUN:( 17.69 ±0.99)mmol/L vs (8.37 ±1.12)mmol/L,t =-23.55,P ＜0.01; ( 12.26 ± 1.11 ) mmol/L vs (8.37 ±1.12 )mmol/L,t =- 19.09,P ＜ 0.01 ;Cr:( 103.22 ± 13.42)μmol/L vs (71.48 ± 8.59) μ mol/L,t =-21.45,P ＜0.01;(86.51 ± 11.49) μmol/L vs (71.48 ±8.59) μmol/L,t =-9.87,P ＜0.01 ;TNF-α:(0.51 ±0.07)ng/ml vs (0.43 ±0.00)ng/ml,t =-5.79,P ＜0.01;(0.47 ±0.03)ng/ml vs (0.43 ±0.00)ng/ml,t =-8.86,P ＜0.01 ].BUN concentration and Cr content and the level of TNF-α in Isoflurane preconditioning group were significantly lower than in I/R group [ BUN:( 12.26 ± 1.1 1 ) mmol/L vs ( 17.69 ± 0.99 ) mmol/L,t =15.67,P ＜ 0.01 ; Cr:( 86.51 ± 11.49) μmol/L vs ( 103.22 ± 13.42 ) μ mol/L,t =6.68,P ＜0.01 ;TNF-α:(0.47 ±0.03) ng/ml vs (0.51 ±0.07) ng/ml,t =2.61,P ＜0.05].Therenal I/R injury which located around kidney tubules was increased in I/R group and isoflurane precondi-tioning group compared to shame operation group [ ( 17.26 ± 1.45 ) vs (0.00 ± 0.00 ),t =- 72.38,P ＜0.01;(12.69±1.83) vs (0.00 ±0.00),t =-39.53,P ＜0.01].The renal I/R injury which located around kidney tubules was decreased in isoflurane preconditioning group compared to I/R group [ ( 12.69 ±1.83) vs (17.26±1.45),t =19.87,P ＜0.01].Conclusions Preconditioning with 1.5％ isoflurane 30 min can protect kidney from I/R injury in rats by regulating the level of TNF-α in renal tissues.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

OBJECTIVETo investigate miR- 125b regulation mechanism by identifying miR-125b target genes and its function in acute myeloid leukemia (AML).

METHODSThe bioinformatics software and database were applied to predict and analyze target genes of miR-125b. The vector contained the target gene 3'-UTR portion cloned into a luciferase reporter construct. A luciferase reporter assay was performed following co-transfection of small molecular miR-125b mimics and target gene wild-type or mutant plasmid into HEK-293T cells. Further in leukemia cell lines NB4 and HL-60, the protein level of target gene was measured by Western blot after overexpression miR-125b. Finally, the viabilities of NB4 and HL-60 cells were measured by CCK-8 assay at 24 h, 48 h, 72 h, 96 h after electroporation.

RESULTSBcl-2-antagonist/killer 1 (Bak1), a pro-apoptotic gene, was a target gene of miR-125b by software predicts. Reporter vector containing the 3'-UTR Bak1 wild and mutation sites were co-transfected with small molecule analogues of miR-125b in HEK-293T cells. Dual luciferase reporter gene assay system showed that miR-125b significantly suppresses the reporter gene activity containing Bak1 3'-UTR by about 53.8% (P<0.05), but it didn't suppresses the reporter gene activity containing 3'-UTR Bak1 mutation. Western blot showed that miR-125b mimics significantly down-regulated the expression of Bak1 in human leukemia cell lines NB4 and HL-60. Meanwhile, the growth rate of cells treated with miR-125b obviously increased compared with that in control by CCK-8 test (P<0.05).

CONCLUSIONOur findings strongly indicated that BAK1 was a downstream target gene of miR-125b, and miR-125b promoted proliferation in human AML cells at least partially by targeting Bak1, so we speculated that miR-125b as an oncogene could be a potential therapeutic target for treating AML.

Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; HEK293 Cells ; HL-60 Cells ; Humans ; MicroRNAs ; genetics ; Transfection ; bcl-2 Homologous Antagonist-Killer Protein ; genetics ; metabolism

OBJECTIVETo observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes.

METHODLymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments.

RESULTThe results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent.

CONCLUSIONThe comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.

Animals ; Calorimetry ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Isatis ; chemistry ; Lymphocytes ; drug effects ; Male ; Mice ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Objective:To investigate the association between choroidal neovascularization (CNV) and the LIPC gene single nucleotide polymorphism (SNP) in a Chinese Han population from Shantou. Methods:A case-control study was designed.Two hundred and twenty-one patients with CNV who visited Shantou International Eye Center from January 2010 to December 2016 were enrolled and served as the CNV group, and 430 healthy volunteers matched in age and gender were enrolled and served as the normal control group.Each of 5 ml fasting peripheral blood of the subjects was extracted, and peripheral blood DNA was extracted after anticoagulation.PCR amplification was conducted on SNP loci of LIPC gene including rs10468017, rs920915 and rs2070895.After purification, genotyping was performed on the above SNP loci using the single base extension (SNaPshot) method.Hardy-weinberg equilibrium (HWE) test was used to determine the genotype frequency of the three SNPs of LIPC gene.The gene frequency and genotype frequency of the 3 loci between the CNV group and normal control group were compared.This study followed the Declaration of Helsinki.Written informed consent was obtained from each subject prior to entering the study cohort.The study protocol was approved by the Ethics Committee of Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong (No.11-004). Results:The genotype frequency distribution of rs10468017, rs920915 and rs2070895 of the three SNPs of LIPC gene reached genetic balance in the total samples ( P>0.05). The genotype frequencies of rs10468017 TT genotype, rs920915 CC genotype and rs2070895 AA genotype in CNV group were 3.62%, 5.43% and 12.22%, respectively, while those of normal control group were 2.56%, 5.58% and 14.19%, respectively, with no statistically significant difference (all at P>0.05). The minimum allele (T) frequency of rs10468017 was 18.1% and 17.2%, the minimum allele (C) frequency of rs920915 was 21.7% and 23.1%, and the minimum allele (A) frequency of rs2070895 was 33.7% and 38.7% in the CNV group and the normal control group, respectively (all at P>0.05). The odd ratio ( OR) values (95%confidence interval [ CI]) of rs10468017, rs920915 and rs2070895 in the CNV group and the normal control group were 1.06 (0.79-1.44), 0.92 (0.70-1.21) and 0.80 (0.63-1.02), respectively. Conclusions:The results from the present study do not indicate the association of LIPC SNPs (rsl0468017, rs920915 and rs2070895) with CNV in the Shantou Han population.

Objective To investigate the clinical value of diffusion tensor imaging (DTI) in the evaluation of cerebral infraction. Methods Sixty-nine patients with cerebral infraction confirmed by clinical manifestation and routine MRI and/or CT were analyzed for the signal intensity changes on DTI. The isotropic apparent diffusion coefficient (ADCiso) and fractional anisotropy (FA) of the infracted area were measured and compared with those of the unaffected side. Results Four types of signal intensity changes on DTI were identified. Type Ⅰ changes were found in 8 infraction lesions, where the ADCiso decreased and FA increased, and the infraction lesions showed hypointensity on ADCiso map and hyperintensity on FA map. Type Ⅱ changes, found in 23 lesions, were characterized by decreased ADCiso and FA values, but ADCiso in the peripheral of the lesions decreased and FA increased, and the lesions were shown as isointensity or hypointensity on ADCiso and FA maps with hyperintensity on the peripheral. Type Ⅲ changes (7 lesions) were manifested by decreased ADCiso and FA values and hypointensity on ADCiso and FA maps. Type Ⅳ changes were found in 31 infraction lesions, showing increased ADCiso and decreased FA with corresponding hyperintensity on ADCiso map and hypointensity on FA map. Significant differences were found in the DTI parameters between the infraction lesions and unaffected side (P<0.05). Conclusion DTI for qualitative and quantitative analysis of cerebral infraction better reveals the pathophysiology of the infraction, allows more precise imaging-based staging of the lesion, and provides evidences for more objective diagnosis, treatment, monitoring and prognostic evaluation of the condition.

Objective:To discuss the application of the classification system for European medical and health care network in the drug related problems in oncology department through the practice of the classification system in clinical cases. Methods:The classifi-cation system (version 8. 0) was used to perform the pharmaceutical care for two patients in oncology department, and the drug related problems ( DRPs) including types, causes, interventions, acceptance degree of interventions and the situation of DRPs were analyzed. Results:DRPs were clearly analyzed by the classification system, which provided scientific and systematic pharmaceutical care for the patients. Clinical pharmacists could find DRPs and provide interventions for the patients, which improved the rationality and safety of medication. Conclusion:Clinical pharmacists establish a perfect pharmacy monitoring system for European medical and health care network, and DRPs can be analyzed by the classification system, which promotes reasonable medication and ensures patients' safety.

OBJECTIVETo probe into the objectivity and authenticity of four properties beteen Mahuang decoction and Maxing Shigan decoction from biothermodynamics.

METHODThe power-time curves of growth of Staphylococcus aureus at different concentrations between Mahuang decoction and Maxing Shigan decoction were determined by TAM Air Isothermal Calorimeter. The growth rate constants of promotive and inhibitory actions were calculated. Moreover, the difference of properties beteen Mahuang decoction and Maxing Shigan decoction was analyzed from the point of view of TCM theory.

RESULTBoth the Mahuang decoction and Maxing Shigan decoction could inhibit the growth and metabolism of Staphylococcus aureus. The k and Pm were decreased with the mass increase of the decoction. However, inhibitory activity of Mahuang decoction with warm property was weaker than that of Maxing Shigan decoction with cool property. Moreover, Mahuang decoction decreased heat output in growth metabolism more weakly than Maxing Shigan decoction. There was a stable difference between them.

CONCLUSIONStudying on biothermodynamics can show the difference of four properties of Traditional Chinese Medicine. So, it provides a new and useful means for the study of the properties of traditional Chinese medicine.

Calcium Sulfate ; chemistry ; Calorimetry ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Ephedra sinica ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Materia Medica ; chemistry ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Prunus ; chemistry ; Staphylococcus aureus ; drug effects ; growth & development ; metabolism ; Thermodynamics

Objective:To evaluate the insulin resistance of patients with polycystic ovary syndrome (PCOS) by hyperinsulin-euglycemic clamp test, and to explore the characteristics of glycolipid metabolism and sex hormone levels in PCOS patients with insulin resistance.Methods:Seventy-three patients with PCOS and 27 healthy women with body mass index and age matched with PCOS patients who were admitted to the Department of Endocrinology of Affiliated Hospital of Zunyi Medical University from July 2017 to February 2019 were underwent hyperinsulin-euglycemic clamp test. All subjects were grouped according to glucose metabolic rate, body mass index, and homeostasis model assessment for insulin resistance (HOMA-IR), the changes and differences of glucose and lipid metabolism and sex hormone indexes in PCOS patients were analyzed.Results:In the PCOS group, impaired glucose regulation accounted for 3.23% (1/31), and abnormal lipid metabolism for 9.68% (3/31). In the PCOS with insulin resistance group, impaired glucose regulation accounted for 7.14% (3/42). Abnormal blood lipid metabolism reached 47.62% (20/42), and 5 patients were diagnosed with metabolic syndrome, accounting for 11.90%. Correlation analysis showed glucose metabolic rate and body mass index, waist-to-hip ratio, systolic blood pressure, fasting plasma glucose, fasting insulin, HOMA-IR, cortisol, triglyceride, total cholesterol, low density lipoprotein-cholesterol (LDL-C), free androgen index (FAI), and glutamyl transpeptidase (GGT) were negatively correlated(all P<0.05), while positively correlated with high density lipoprotein-cholesterol (HDL-C; P=0.028). HOMA-IR was positively correlated with body mass index, waist-to-hip ratio, systolic blood pressure, fasting plasma glucose, fasting insulin, HbA 1C, LDL-C ( P<0.05), and negatively correlated with glucose metabolic rate and HDL-C ( P<0.05). Body mass index and waist-to-hip ratio, systolic blood pressure, fasting plasma glucose, fasting insulin, HOMA-IR, triglyceride, and LDL-C ( P<0.05) were positively correlated, and negatively correlated with glucose metabolic rate, HDL-C, and sex hormone binding globulin (SHBG; P<0.01). Multivariate linear stepwise regression analysis showed that body mass index, total cholesterol, triglyceride, and cortisol were principal factors affecting glucose metabolic rate. Fasting plasma glucose, fasting insulin, and systolic blood pressure were important factors influencing HOMA-IR. Glucose metabolic rate, HOMA-IR, HDL-C, while SHBG were still vital to body mass index. Conclusion:FAI, SHBG, and cortisol may be involved in the insulin resistance development of PCOS patients, and PCOS patients with insulin resistance were more susceptible to metabolic disorders.

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult