To offer the guidance of the registration application of in vitro diagnostic (IVD) reagents,the technical requirements for registration of the pathogen-specific Immunoglobulin M (IgM) qualitative test reagents were discussed.The application paperwork including the evaluation of analytical performance was analyzed based on the the legal requirement from the China Food and Drug Administration (CFDA)and the experience of the IVD registration.

BACKGROUND:Currently, there are many problems existing in the construction of large tissue-engineered bone, and the most important is ischemic necrosis. Vascularization is critical for the biological function of large tissue-engineered bone.
OBJECTIVE:To prepare large pieces of vascularized tissue-engineered bone for orthotopic transplantation in repairing large femoral defects of rabbits, and to explore tissue regeneration methods and characteristics.
METHODS:Rabbit femoral structure was firstly imitated to construct a vascularized prefabricated model, based on which, we constructed large pieces of vascularized tissue-engineered bone. A total of 18 New Zealand white rabbit 18, aged 8 weeks, were divided into experimental group and control group. The experimental group was subject to orthotopic transplantation of large vascularized tissue-engineered bone, while the control group was treated with orthotopic transplantation of large tissue-engineered bone without vascularization processing. After 2, 4, 8 weeks, gross observation, X-ray films, and tissue section were compared between the two groups.
RESULTS AND CONCLUSION:Gross observation and imaging observation showed that the osteogenesis in the experimental group at 2, 4, 8 weeks were better than that in the control group. The ratio of new bone tissue to total area of bone graft was significantly higher in the experimental group than the control group at 2, 4, 8 weeks postoperatively (P<0.01). These findings indicate that large vascularized tissue-engineered bone is preferred to the repair of large bone defects.

POCT in vitro diagnostic reagents featuring with easy operation , on-site testing, low requirements for specimen processing , fast results , usually applied small testing equipment , is a strong complement to high-throughput , large automated tests in clinical laboratory centers.According to the State Council issued in 2014 “Supervision and Regulation of Medical Devices ” and series regulations and management practices issued by the China Food and Drug Administration , conduct an investigation in POCT IVD registration relevant technical requirements and give some advice for the relevant personnel of register work.

Objective To establish an anodic-stripping voltammetry method for determination of trace cadmium in water using dithizone-modified glassy carbon electrode. Methods Cd2+ was preconcentrated on the surface of the dithizone-modified glassy carbon electrode at -1.21 V, and subsequently reduced to Cd on the electrode. When the electric potential of electrode increased from -1.21 V to -0.30 V, the reduced cadmium was oxidized and stripped from surface of electrode, and the sensitive anodic-stripping peak was observed at -0.79 V. The parameters including the selection of supporting electrolytes, pH value, the amount of dithizone, concentration electric potential and time were optimized. Results The most suitable reaction medium was 0.1 mol/L NaAc-Hac-KI solution at pH value of 3.00.The most suitable volume of 1g/L methenyl chloride-dithizone solution was 10 ?l. The optimized concentration electric potential and time were -1.21 V and 5 min respectively. The linear range of this method was 5?10-8-2?10-6 mol/L. The average recovery rate was 98.42%. The relative standard deviation was 2.90%(n=8). The detection limit was 6?10-9 mol/L. Conclusion The method was simple ,rapid and sensitive,which was suitable for the determination of trace cadmium in practical water sample.

Diagnostic reagents of polymerase chain reaction ( PCR ) nucleic acid amplification is growing more and more important in the field of clinical diagnosis along with the development of the PCR technology.During to the complexity of the detection target and the clinical requirements for measuring linearity, accuracy, specificity, subtype detection, precision, limit of detection/quantification and interfering substance , the diagnostic reagents of nucleic acid amplification have a very strict control system . In order to ensure the quality of the experimental data of quantitative PCR , there have been standards such as The Minimum Information for Publication of Quantitative Real-Time PCR Experiments ( MIQE) , aiming to putting forward specific requirements for sample processing , experimental designing , system setting and quality control.But in the clinical application of nucleic acid amplification products , lots of requirements are not clear for the PCR system setup .This article discussed the requirements of PCR system setting , quality control system and Methodology of nucleic acid extraction and purification , aiming to promote the related design and the establishment of the standards of relevant .

BACKGROUND: Traumatic brain injury generates a cascade of arachidonic acid metabolic events that mainly presented by the increment of prostaglandin and oxygen free radicals. Indomethacin can potently inhibit the activity of cyclooxygenase, decrease the synthesis of prostaglandins, and may decrease the production of oxygen free radical, and thus may attenuate the pathological changes of brain injury.OBJECTIVE: To observe the changes of prostaglandin in early brain injury and after indomethacin intervention, so as to explore the pharmacological mechanism of indomethacin.DESIGN: A randomized and controlled trial based on experimental animals.SETTING: Department of neurosurgery and department of cerebral surgery in a university hospital.MATERIALS: This study was carried out at the Laboratory of Neurosurgery Department, Medical College of Southeast University between March and September 2000. Thirty-six hybrid cats were randomly divided into normal control group, brain injury group and indometbacin intervention group, with 12 cats in each group.INTERVENTIONS: Brain injury was simulated according to previously reported grading mechanical traumatic animal model establishment; cats with medium brain injury were enrolled in this study. The ultimate concentrations of prostacyclin (PGI2) and thromboxane A (TXA2) to 6-keto-prostaglandin F 1 alpha(6-keto-PGF1α) and thromboxane B2(TXB2) in brain vein blood, as well as total brain superoxide dismutase(SOD) and cerebral water content were measured 6 hours after trauma.MAIN OUTCOME MEASURES: 6-keto-PGF1α, TXB2, SOD, and cerebral water content.RESULTS: Both 6-keto-PGF1α and TXB2 in brain vein blood remarkably increased in early brain injury[from(0.057±0.010) g/L to (0.264±0. 126) g/L, from(0. 060 ±0. 012) g/L to(0. 134 ±0. 048) g/L respectively], with the increment of the former higher than the latter, the ratio of TXB2/6-keto-PGF1α decreased from 1. 052 ±0. 145 to 0. 545 ±0. 184, and cerebral water content increased from(77.39 ± 0. 36)% to (78.06±0.41)% ; meanwhile, total brain SOD significantly decreased from (94. 869 ± 5. 418) μkat/g to(54. 368 ± 3. 417) μ kat/g( P ＜ 0.01) . In contrast to brain injury group, the concentrations of 6-keto-PGF1α and TXB2 in indomethacin intervention group significantly decreased, which were similar to those of control group, but the total SOD significantly increased from (54. 368 ±3. 417) pkat/g to (81. 433 ±7. 268) μkat/g (P ＜0. 01), and water content lightly decreased without statistical significance( P ＞ 0. 1 ).CONCLUSION: PGI2 and TXA2 increase in early brain injury in experimental cat model, accompanied by free radical synthesis, resulting in the exacerbation of brain injury. Indomethacin may be helpful to relieve posttraumatic secondary brain injury by regulating the imbalance of PGT2 / TXA2 and decreasing the production of free radical.

Objective The present study was to analyze the role of ACh-lateral septum(SL)pressor system in the Central amygdaloid nucleus(AC)-emotional pressor circuit.Method Interurban microinjection of different drugs,then blood pressure and heart vate were vecorded.Results(1)CRF(corticotropin releasing factor)or SP(substance P)microinjection into the SL can induced pressor responses.The AC pressor responses to glutamate(Glu)could be attenuated by preinjection of CRF antagonist or SP antagonist into bilateral SL.(2)the AC pressor responses to Glu were also reduced by preinjection of atropine into either bilateral HBL,LC or RVL respectively,(3)Since lateral HB(HBL)projects to the posterior hypothalamus(HP)containing ACh-ergic neurons,and excitation of the latter produces pressor response via the RVL and LC-RVL;atropine preinjection into the HP could also decrease the HBL-and AC-pressor response.Conclusion The present results indicate that the AC produce pressor responses involved with the SL via their CRF and SP,mechanism underlying pressor response of SL is involved in pressor response of AC.

Objective To establish a new post-embedding immunoelectron microscopy method for localization of protein antigens in central nervous system using colloidal golds as probes.Methods Rat brain tissue sections were fixed with osmium-ferrocyanide mixture.Free floating frozen sections were collected to determine the retrievability of antigens by sodium metaperiodate(NaIO_(4)).Then the best concentration of NaIO_(4) for maximal antigen retrieval was determined on hydrophilic Technovit 7100 resin-embedded semithin sections.Finally,the corresponding antigen was labeled with colloidal golds on ultrathin sections.Results NaIO_(4) showed different retrieving effects on different antigens;to those antigens that could be effectively retrieved by NaIO_(4),low concentration of NaIO_(4)could realize antigen retrieval;EM microscopy showed that ultrathin sections treated by low concentration of NaIO_(4) allowed higher density of colloidal gold labeling without apparently compromising the ultrastructures of tissues.

Objective To investigate the effects of Huangkui Capsules on oxidative stress and micro-inflammation state in diabetic nephropathy patients with maintenance hemodialysis. Methods Forty-six patients with diabetic nephropathy who had received maintenance hemodialysis for over six month in Gansu Provincial People's Hospital were randomly divided into observation group (23 cases) and treatment group (23 cases) from January to December in 2014. Twenty healthy subjects who had physical examinations were selected as the control group at the same time. Observation group were given conventional hemodialysis for 4 hours, 3 times a week; treatment group were given conventional hemodialysis combined with Huangkui Capsules, orally, 2.5 g each time, 3 times a day. Both groups were treated for 8 weeks. Blood was collected before and after treatment for detection of hypersensitive C-reactive protein (hs-CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), white blood cell (WBC), malondialdehyde (MDA), cycle of advanced oxidation protein products (AOPP), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. Results The levels of IL-6, TNF-α, AOPP, WBC, MDA and hs-CRP were significantly higher in the observation group and the treatment group compared with the control group before treatment (P<0.01), SOD and GSH-Px levels decreased (P<0.01); there was no statistical significance between the observation group and the treatment group (P>0.05). After treatment, the levels of IL-6, hs-CRP, MDA and AOPP increased in the observation group (P<0.05), GSH-Px and SOD decreased (P<0.05), there were no statistical significance in TNF-αand WBC (P>0.05);The levels of AOPP, TNF-α, hs-CRP, WBC, MDA and IL-6 in treatment group significantly decreased (P<0.01), GSH-Px and SOD significantly increased (P<0.01); there was statistical significance between the observation group and the treatment group (P<0.01). Conclusion Micro-inflammation state and oxidative stress exist generally in diabetic nephropathy maintenance hemodialysis patients. Huangkui Capsules can improve oxidative stress and micro-inflammation state in diabetic nephropathy maintenance hemodialysis patients.

Objective To observe the change of tumor cells on sodium phenylacetate(NaPA) treating G422 glioma mice,and explore its mechanism.Methods Two kinds of mice models(intracranial and muscular G422 glioblasloma cells) were established,and were divided into five groups,among which 3 groups for expe rimental groups,they were received NaPA(1200,800,400 mg/kg?d).In the other two groups,the positive control group was administered BCNU(20 mg/kg) only one time.The negative control group was given saline 24 h later tumor inoculation,these mice bagan to be administered.The experimental group and negative control group were injected NaPA and saline for 14 days respectively.After that,the drug toxicity,the survival period,survival rate of the mice were observed,and the pathology and ultrastructure of glioma in every group were also observed.The muscular tumor mice were sacrificed for measurement of tumor suppression rate.Results NaPA can prolong the life span of the mice with glioma;it has concentration dependent inhibition to glioma proliferation;the pathology and electron microscopy showed the decrease of glioma nuclei fission image treated by NaPA,the increase of rough endoplasmic reliculum and the cell proptosis were found,it demonstrated the tumor cells had differentiation trend.Conclusion NaPA had antitumor effect by inducing glioma cell differentiation and inhibiting the growth of tumor.