Objective To establish a simple, cost-effective method to measure reactive oxygen metabolites ( ROMs) using automatic biochemical analyzer.Methods Serum samples were collected from 50 healthy individuals and 50 hospitalized patientsin Beijing Hospital from September 2013 to November 2013.By setting up and optimizing parameters on automatic biochemical analyzer, a new method of measuring ROMS was established with TBHP as calibrator.130 healthy non-smokers were recruited in Beijing Hospital from November 2013 to December 2013 to establish reference interval, including 73 males and 57 females.The average age was 68.8 ±12.5.According to CLSI documents, there was the establishment of the new method including precision, limit of blank ( LoB ) , limit of detection ( LoD ) , limit of quantitation (LoQ), linearity,interference testing, and reference interval.Results The intra assay imprecision was 1. 5%-4.1% and the total imprecision was 4.4%-9.9%.LoB was 0.85 mmol/L TBHP;LoD was 2. 7mmol/L TBHP;LoQ was 5.9mmol/L TBHP.The linearity was 5.9-600 mmol/L (R2 =0.995).When triglyceride≤28.58 mmol/L, hemoglobin≤173 g/L, vitamin C≤60 mg/dl and bilirubin≤73.8 μmol/L, the deviation was -7.6%, 4.6%, -98.9%, 19.1%respectively.The normal reference interval was 61.9 -88.1 mmol/L TBHP.Conclusions The newly-established method has good performances of precision,LOD and linearity, which can meet the clinical needs.The interference of triglyceride and hemoglobin is small, while vitamin C and bilirubin have stronger interference on measurement.( Chin J Lab Med,2015,38:163-167)

ObjectiveTo detect specific antigens of neural cells in amniotic epithelial cells(AECs) in rats. MethodsAECs were dissociated and purified from the amnion of pregnancy 12—14 d rats. The expression of specific markers of neural stem cells (Nestin, Musashi) and differentiated cells (MAP-2,NSE,GFAP) and ChAT, NT-3 in the AECs were detected by immunocytochemistry. ResultsThe cultured AECs displayed positive immunoreactivity to MAP-2, NSE, GFAP, Nestin and Musashi. In addition, the cells also demonstrated immunoreactivity to ChAT and NT-3. ConclusionAECs are similar with neural cells and it may be useful as a sustained source to improve outcome of neural stem cells transplantation.

ObjectiveTo investigate an effective method to isolate neural stem cells(NSCs).MethodsNSCs were dissociated by digestion with trypsin, EDTA and different doses of Dispase,and serum-free culture techniques and immunohistochemistry techniques were used to verifying the dissociated.ResultsA lot of single neural stem cells were obtained by using Dispase to digest neurosphere, and the cells could keep its structure and morphology.ConclusionIt is an ideal method by using Dispase to digest neurosphere for isolating NSCs.

Objective To investigate the alterations of peripheral blood T lymphocyte subsets in patients with sepsis and septic acute kidney injury, and explore the clinical significance.Methods Fifty-five patients with sepsis and forty-three patients with septic acute kidney injury were enrolled in this study. At the same period, thirty healthy subjects were enrolled as the control group.T lymphocyte subsets inclu-ding CD3 +T, CD4 +T, CD8 +T cells, and CD4 +T/CD8 +T in peripheral blood were detected by flow cy-tometry, and acute physiology and chronic health evaluation Ⅱ( APACHE Ⅱ) were graded within twenty-four hours after admission.Then, correlation of the APACHEⅡscores and T lymphocyte subsets was ana-lyzed.Results In the septic acute kidney injury group, peripheral blood CD3 +T, CD4 +T cell percenta-ges, and CD4 +T/CD8 +T ratio were significantly lower than those in the control group and the sepsis group (all P <0.05).In the septic acute kidney injury group with stage 3, CD3+T, CD4 +T cell percentages, and CD4 +T/CD8 +T ratio in the patients were significantly lower than those in stage 1 and stage 2 ( all P <0.05).In the septic acute kidney injury group,CD3 +T, CD4 +T cell percentages, and CD4 +T/CD8 +T ra-tio were significantly lower in dead group than those in survival group (all P <0.05).APACHEⅡscores in patients with sepsis were significantly negatively correlated with peripheral blood CD4 +T cell percentages and CD4 +T/CD8 +T ratio ( r =-0.645,-0.492, allP <0.05).Conclusions There are varying de-grees of cellular immune imbalance in patients with sepsis and septic acute kidney injury, characterized by decline of circulating CD3 +T, CD4 +T cell percentages, and CD4 +T/CD8 +T ratio.CD4 +T cell percenta-ges and CD4 +T/CD8 +T ratio are closely related to the severity and prognosis of septic acute kidney injury.

Purpose To investigate the value of MRI T2 mapping imaging in diagnosis of lumbar intervertebral disc degenerative (IVDD).Materials and Methods A prospective study was performed on 100 patients who underwent 1.5T MRI examination because of lumbar and back pain from October 2013 to December 2015 at the Fifth People's Hospital of Shanghai,Fudan University.The MRI examinations included conventional sagittal T1WI and T2WI,axial T2WI and median sagittal T2 mapping imaging.According to Pfirrmann standard,475 lumbar intervertebral discs were graded.T2 values of nucleus pulposus (NP) and annulus fibrosus (AF) of anteriority and posterior with different grades were measured and compared.The correlations between T2 value,age and grade were further analyzed.30 patients were selected and underwent a second MR examination after half a year,and the difference of T2 values between the two MR examinations were compared.Results Except for grade Ⅳ and grade Ⅴ,the differences of T2 values of NP between the other grades of lumbar intervertebral discs were statistically significant (P＜0.05).T2 values of NP were negatively correlated with the grades (r=-0.77).There were no significant differences in T2 values of NP and AF of anteriority and posterior in 30 patients with back pain between the two MR measurements of half year interval (P＞0.05).Conclusion T2 mapping imaging can quantitatively assess the degree of IVDD,especially T2 values of NP can reflect the differences of IVDD with grade Ⅰ-Ⅳ,so it can provide the imaging evidence for the diagnosis of IVDD.

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end ofa 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall ofNHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

In recent years, polysaccharides have received much attention because of their high safety and good immunological activity. The study of polysaccharide in vivo process is a key scientific problem that needs to be solved for polysaccharide drug development. Some progress has been made in the field of polysaccharide pharmacokinetics and immunomodulation. However, due to the lack of both chromogenic and light-absorbing groups and the complex molecular structure of polysaccharides, the in vivo processes and immunomodulatory mechanisms of polysaccharides have been slow to be investigated. The effective combination of multiple techniques can break the bottleneck of difficult tracing and unknown immunomodulatory mechanism of polysaccharides in vivo, and promote the development and utilization of polysaccharides. In this paper, we systematically summarize the key techniques in the study of polysaccharide in vivo processes and immunomodulatory mechanisms in order to provide technical references and research ideas for the study of polysaccharide in vivo processes and immunomodulatory mechanisms.

Along with the development of economy and society, type 2 diabetic mellitus (T2DM) has become one of the most common diseases at the global level. As one of the complications of T2DM, diabetic neuropathic pain (DNP) stubbornly and chronically affects the health and life of human beings. In the pain field, dorsal root ganglion (DRG) is generally considered as the first stage of the sensory pathway where the hyperexcitability of injured neurons is associated with different kinds of peripheral neuropathic pains. The abnormal electrophysiology is mainly due to the changed properties of voltage-gated sodium channels (VGSCs) and the increased sodium currents (I(Na)). Curcumin is an active ingredient extracted from turmeric and has been demonstrated to ameliorate T2DM and its various complications including DNP effectively. The present study demonstrates that the I(Na) of small-sized DRG neurons are significantly increased with the abnormal electrophysiological characteristics of VGSCs in type 2 diabetic neuropathic pain rats. And these abnormalities can be ameliorated efficaciously by a period of treatment with curcumin.
Animals ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Neuropathies ; drug therapy ; Ganglia, Spinal ; cytology ; drug effects ; metabolism ; Neuralgia ; drug therapy ; Neurons ; drug effects ; metabolism ; Rats ; Sodium ; Voltage-Gated Sodium Channels ; physiology

To determine the optimum process conditions for dry granulating technique of Qibai Pingfei granule, granule excipient type, rolling wheel speed and pressure and feeding speed were studied. Taking shaping rate at a time, moisture absorption and dissolubility as index, the type and amount of granule excipient were determined. In addition, taking shaping rate at a time as index, parameters of rolling wheel speed and pressure and feeding speed were researched through single factor test and response surface methodology. The optimum parameters were as follows: lactose as excipient, dry extract powder to excipient at 1:2, rolling wheel speed and pressure at 10.9 Hz and 6.4 MPa and feeding speed at 7.2 Hz. After validation of three batches pilot-scale production, the optimum processing parameters for dry granulating technique of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
Drugs, Chinese Herbal ; Powders ; Technology, Pharmaceutical ; methods

BACKGROUND: Ulinastatin (UTI) is a urinary trypsin inhibitor extracted and purified from urine of males. This study aimed to explore the effects of UTI on paraquat-induced-oxidative stress in human type II alveolar epithelial cells. METHODS: The human type II alveolar epithelial cel s, A549 cel s, were cultured in vitro. The A549 cel s were treated with different concentrations of paraquat (200, 400, 600, 800, 1000, 1200 μmol/L) and ulinastatin(0, 2000, 4000, 6000, 8000 U/mL) for 24 hours, the cell viability was measured by cell counting kit-8 and the median lethal concentration was selected. In order to establish an in vitro model of paraquat intoxication and to determine the safe dose of ulinastatin, we calculated LD50 using cell counting kit-8 to determine the survival rate of the cells. A549 cells were divided into normal control group, paraquat group and paraquat+ulinastatin group. The levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were detected by biochemistry colorimetry, while the level of reactive oxygen spies (ROS) was detected by DCFH-DA assay. RESULTS: The survival rate of A549 cells treated with different concentrations of paraquat decreased in a concentration-dependent manner. Whereas there was no decrease in the survival rate of cells treated with 0–4000 U/mL ulinastatin. The levels of MDA, MPO, and ROS were significantly higher in the paraquat group than in the normal control group after 24-hour-exposure. And the survival rate of the paraquat+ulinastatin group was higher than that of the paraquat group, but lower than that of the normal control group. The levels of MDA, MPO, and ROS were lower than those of the paraquat group. CONCLUSION: Ulinastatin can alleviate the paraquat-induced A549 cell damage by reducing oxidative stress.