Polymerase chain reaction (PCR) is one of the common techniques in molecular biology, which can amplify nucleic acids through the cycle of denaturation, annealing and extension. Based on the principle of common PCR, rapid PCR is to realize the amplification of nucleic acids in less time without affecting the specificity, sensitivity and fidelity of the reaction. A lot of research work in this field has been going on in recent years. This article will make a review of the development of rapid PCR with emphases on the improvement of DNA polymerase, the choice of additives and the improvement of thermocyclers.

Objective To establish virtual three-dimensional instrument and cerebral aneurysm models by using three-dimensional moulding software,and to explore the effect of the models in interventional preoperative simulation.Methods The virtual individual models including cerebral arteries and aneurysms were established by using the three-dimensional moulding software of 3D Studio MAX R3 based on standard virtual cerebral aneurysm models and individual DSA image.The virtual catheter,guide wire,stent and coil were also established.The study of interventional preoperative simulation was run in personal computer,and included 3 clinical cases.Results The simulation results of the working angle and the moulding angle of the head of catheter and guide wire in 3 cases were identical with that of operation results. The simulation results of the requirement of number and size of coil in 1 case of anterior communicating aneurysm and 1 case of posterior communicating aneurysm were identical with that of operation results.The simulation results of coil for aneurysmal shape in 1 case of giant internal carotid artery aneurysm were more than 2 three-dimensional coils with size of 3mm?3 cm from the operation results,and the position of the second coil in aneurysmal neck was adjusted according to the results of real-time simulation.The results of retrospective simulation of operation procedure indicated that the simulation methods for regular and small aneurysms could become a routine simulation means but more simulation experience was needed to build up for the giant aneurysms.Conclusions The virtual three-dimensional instrument and cerebral aneurysm models established by the general software provided a new study method for neuro-interventional preoperative simulation,and it played an important guidance role in developing neuro- interventional operation.

OBJECTIVE:To observe the occurrence of paclitaxel(PTX)-induced muscle soreness and therapeutic efficacy and safety of diclofenac sodium. METHODS:Among 84 patients with malignant tumor receiving PTX chemotherapy,56 patients suf-fered from PTX-induced muscle soreness,among which 22 female patients suffered from medium and severe muscle soreness and then were randomly divided into group A and B,with 11 cases in each group. Group A was given Diclofenac sodium sustained-re-lease tablet 75 mg orally,once a day;group B was given Paracetamol and tramadol hydrochloride tablets one tablet orally,once a day,and then given Promethazine hydrochloride injection 100 mg subcutaneously,2-3 times a day when muscle soreness could not be born. Treatment course of both groups lasted for 5 d. The distribution of muscle soreness were observed. The onset time and dura-tion of muscle soreness were also observed as well as pain relief and the occurrence of ADR in group A and B. RESULTS:Among 84 cases,the incidence of muscle soreness was 66.67%,among which mild pain accounted for 23.81%,moderate pain accounted for 13.10%,and severe pain accounted for 29.76%. Among 56 patients with muscle soreness,earliest muscle soreness occurred on the day of medication,and most of muscle soreness occurred on 1-3 days after medication,mainly manifesting as sore,activity limitation when severe,associated with fatigue. The pain relief rate of group A and B were 100%,but the incidence of adverse re-actions in group A was significantly lower than group B,with statistical significances(P<0.05).CONCLUSIONS:Diclofenac sodi-um is similar to paracetamol and tramadol hydrochloride in the treatment of PTX-induced muscle soreness,but it is better than paracetamol and tramadol hydrochloride in safety.

Background It is thought in recently that photodynamic therapy (PDT) is an effective treatment method for chronic central serous chorioretinopathy (CSC), but the dosage of verteporfin and its long-term efficacy and complications is rarely elucidated ever before.Objective This study was to observe the long-term efficacy and safety of 60％ dose verteporfin PDT for chronic CSC.Methods This is a retrospective study and a self-controlled design was used.The clinical data of 25 eyes of 21 chronic CSC patients who received 60％-dose verteporfin PDT in Henan Eye Institute from January 2009 to May 2010 were reviewed, with the male 18 (85.71％) and female 3 (14.29％) as well as monocular CSC 17 patients and binocular CSC 4 patients.The average ages of the patients were (43±5) years.Fundus fluorescein angiography (FFA) , indocyanine green angiography (ICGA), optical coherence tomography(OCT) and best corrected visual acuity (BCVA) were examined in all the patients before and after treatment.PDT with the 60％-dose verteporfin (3.6 mg/m2) was carried out on the CSC eyes.The treated eyes were examined 2 weeks, 1 month and 3 months after PDT.The BCVA,subfoveal choroid thickness,FFA and ICGA findings before and after PDT were compared.The following-up duration was 5 years or more.Results The BCVA before and 3 months after PDT were 0.5 ±0.1 and 0.9±0.2, respectively, with a statistically significant difference between them (t =19.17,P =0.00).The subfoveal choroidal thickness value 3 months after PDT was (326.56±39.47) μm,which was significantly reduced in comparison with (486.24 ±47.53) μm before PDT (t =25.17, P =0.00).FFA and ICGA showed that the leakage of fluorescein (hyperfluorescence) was disappeared in all the treated eyes.No systemic or local adverse effects and recurrence were observed during the follow-up period.Conclusions On the basis of the results of this study and available information,60％-dose verteporfin PDT seems to have a better long-term efficacy and safety than full-dose verteporfin in treating chronic CSC.

Nitric oxide (NO), which is involved in the regulation of the cardiovascular system, nervous system, immune system, reproductive system, digestive system and other physiological activities, is an important biological substance with activity. Under normal physiological conditions, neuronal nitric oxide synthase (nNOS) can precisely regulate the nervous system NO production, release, diffusion and inactivation processes. But an excess of NO associates with the development of cerebral ischemia, Alzheimer's and Parkinson's psychosis nervous system diseases, while inhibition of nNOS activity can regulate the content of NO in vivo, and produce a therapeutic effect on some of the nervous system diseases. This review mainly describes the structure and regulation of nNOS and recent developments of small molecule inhibitors of nNOS.
Alzheimer Disease ; physiopathology ; Brain Ischemia ; physiopathology ; Humans ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type I ; antagonists & inhibitors ; metabolism ; Parkinson Disease ; physiopathology

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

OBJECTIVETo compare the differences between computer aided identification and ultra-thin layer slicing in measuring the lesions of femoral head necrosis,and to confirm the accuracy and practicability of computer aided method.

METHODSFrom June 2012 to December 2013, the X-ray and MRI of 24 patients (24 hips on unilateral) were reviewed, who had avascular necrosis of the femoral head (ANFA) at late stage (stage III and IV) according to the ARCO international staging system. There were 15 males and 9 females, with an average age of (65.1 ± 8.8) years old, ranged 33 to 74 years old. Based on the software system with seeds point identification, the ragional adaptive search method with computer aid was used to calculate the volume of necrotic lesion in femoral on MRI. Then the pathological slices of those intraoperative femoral heads were made to measure the gross volume of necrotic lesion in femoral head,and the values were compared with the data in the computer.

RESULTSFor 24 hips, by the calculation of computer, the necrotic volume was (20.00 ± 3.04) cm (ranged, 18.72 to 21.29 cm³). Under the pathological section, the necrotic volume of the femoral head was (19.89 ± 3.17) cm³ (ranged, 18.55 to 21.23 cm³). In computer and pathology two kinds of measurement, the two entire femoral head volume had no significant difference using these two measurements (t = -1.227, P = 0.232).

CONCLUSIONComputer aided identification for necrotic area of femoral head adaptive can demonstrate the morphology of femoral head necrosis accurately and reliably, which will help surgeon better understand the morphology and orientation in femoral head.

Adult ; Aged ; Diagnosis, Computer-Assisted ; Female ; Femur Head Necrosis ; pathology ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged

Objective To build the experimental basement for the clinical use of cytokines induced killer(CIK)cells from the cord blood mononuclear cells(CBMNC)in tumor adoptive cellular immunotherapy, an effective protocol for their proliferation in vitro and cytotoxicity of CIK cells was established.Methods The lymphocytes from umbilical cord blood were isolated by density gradient centrifugation and suspended in medium with CD_3 mAb,rIL-2,rIL-1 and IFN-? as inducing agents to prepare CIK cells.At the same time, the lymphokine activated killer(LAK)and CBMNC were set as controls,which were only added IL-2 and not any cytokines during the whole culture.The changes of CIK cells before and after induction were observed with microscope and the phenotypes of the cells were analyzed by using flow cytometry.The proliferation of CIK cells were determined by trypan blue exclusion assay and the cytotoxic activity to lung cancer cell were tested with MTF method.Results According to the experiment,combining use of four types of cytokines could generate a great deal of CIK cells possessing highly cytotoxicity.From day 5 CIK cells became to prolif- erate and reached the peak at day 14.During the whole period,the relative percentage of CD_3~+ CD_(56)~+ cells in- creased significantly.Compared with LAK cells,which reached the proliferation peak at day 7 and then showed no evident proliferation.The control cells(CBMNC)showed no evident change of phenotypes and proliferation.CIK cells showed a higher antitumor activity on the tumor cells than LAK cells and CBMNC in vitro.Conclusion Umbilical cord blood can generate a great deal of CIK cells combining used with cy- tokines.Compared with classic LAK cells,umbilical cord blood CIK cells have the advantages of rapid prolif- eration speed and powerful cytotoxicity.CIK cells will be promising as a new strategy for the adoptive cellular immunotherapy of tumor.

Objective To knowthe effect of selenium on expression ofnitric oxide synthase(eNOS) and inducible nitric oxide synthase(iNOS) induced by fluoride in ECV-304(human umbilical vein endothelial cells).Methods The cells,ECV-304,were divided into two groups and treated as follows,one group was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L),the other was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L)and added sodiumselenite(100 nmol/L) respectively.24 h of treatment,The activity ofeNOS and iNOS were measured by NO and NOS assay kits and the expression of eNOS mRNA and iNOS mRNA was analyzed by RT-PCR.Results The activity of eNOS and the expression of eNOS mRNA were down-regulated by fluoride with a dose dependent manner,while the activity of iNOS and the expression of iNOS mRNA were up-regulated.Selenium showed a significant antagonism to low level fluoride treated groups.Conclusion Seleniumcan antagonize the effect offluoride on the expression ofiNOSand eNOS in the vein endothelial cells when the fluoride exposure level is not toohigh.

Parkinson's disease(PD) is a progressive neurodegenerative disease characterized by motor symptoms such as rigidity, rest tremor, and bradykinesia. However, evidence demonstrated that PD encompasses several non-motor disturbances as well, such as cognitive impairment. Mild cognitive impairment can be present since early stages of the disease, and characterized by impairments in several cognitive domains including executive functions, attention, and visuospatial skills, language, and memory; In advanced stages of the disease, cognitive defects can develop into dementia and there is a considerable heterogeneity in the cognitive impairments. In this review, we focus on the clinical characteristics, biomarkers, and drug and non-drug treatments of cognitive impairment in Parkinson's disease, hoping to provide help for clinical practice.