Objective To develop a double antibody sandwich ELISA for the determination of the Coxsackievirus A6(CVA6) antigen content, and to verify and preliminarily apply it for the CV-A6 production process and the quality control of tetravalent hand-foot-mouth disease(HFMD) vaccine.Methods Paired antibodies were screened and double antibody sandwich ELISA was developed by using purified CV-A6 sheep polyclonal antibody as coating antibody and mouse anti-CVA6 monoclonal antibody as detection antibody. The experimental conditions were optimized, and the linearity, range,specificity, accuracy, precision and robustness of the method were verified. The developed method was used to detect the antigen content of CV-A6 antigen production process samples and final products.Results Both of the sheep polyclonal antibody and monoclonal antibody 16D1-1E5 had neutralizing antibody activity, while the other three monoclonal antibodies showed no neutralizing antibody activity. The ELISA titers of all antibodies were not lower than 1. 00 × 105. The monoclonal antibody 2B6-4F9 was IgG3 subtype, while the other three monoclonal antibodies were IgG2a subtype. Using sheep polyclonal antibody as coating antibody and monoclonal antibody 16D1-1E5 as detection antibody for pairing, the method was developed.The dilution ratios of coating antibody and detection antibody were 1∶2 000-1∶6 000 and 1∶4 000-1∶6 000 respectively.The linear range of the developed method was 0. 664-21. 263 U/mL, with the R2≥ 0. 99. The method could only detect CVA6 antigen specifically and had no cross-reaction with CV-A10, CV-A16 and enterovirus 71(EV-71) antigens. M199,DMEM, Vero cell protein and bovine serum protein probably existing in the production process had no interference with this method. The measured/theoretical values of samples with high(21. 00 U/mL), medium(10. 50 U/mL) and low(1. 70 U/mL)concentrations were between 95% and 105%, with the RSDs within 15%. In the range of 0. 664-21. 263 U/mL, the R2was not less than 0. 99, the measured/theoretical values were between 95% and 105%, and the RSDs of precision verification was within 15%. The method had no difference in accuracy and precision at different incubation temperatures(35, 37, 39 ℃).The antigen content of CV-A6 antigen production process samples and finished products detected by the developed method effectively reflected the changing trend of antigen concentration in process samples and final products.Conclusion The double antibody sandwich ELISA for quantitative determination of CV-A6 antigen was developed with good linearity, range,specificity, accuracy, precision, and robustness, which can be used for the quality control of CV-A6 antigen process and final products, and lays a foundation for the in vitro efficacy evaluation of CV-A6 antigen.

In order to sum up the antineoplasmic active components of Holothuria, we studied the literatures in recent years. The results showed that Holothuria contained a variety of anti-tumor active ingredient, such as Holothurian Glycosaminoglycan and Holothurin. These anti-tumor active components played important roles in different stages of tumor developmment, progression and metastasis. The discovery of new anti-tumor active ingredients may be helpful for the application of Holothuria in clinical tumor treatment.

PBL teaching method can cultivate students’ ability to analyze the existing problems, find out more problems and resolve the problems finally, and it is important to improve students’ comprehensive quality. The experimental teaching innovation of Histology and Embryology is summarized in this article, which turned induction, practice and examination, the three stages of experimental teaching into" Problem-Based learning", thus making the teaching effect much better.

Objective To study the radiosensitization of histone deaeetylases inhibitor(HDACI) panobinistat in prostate cancer cells in vitro,as well as the possible mechanisms.Methods IC20 of two prostate cancer cell lines(LNCaP and PC-3)was determined using MTI assay.Cells received a single dose irradiation of 0,2,4,6,or 8 Gy using 6 MV X-ray for radiosensitivity experiment,but only 2 Gy for western blot and flow cytometry.Radiosensitization of panobinostat was investigated with clonogenic assay,and sensitizing enhancement ratio(SER)was calculated with single-hit multi-target model.Western blot was used to compare γH2AX expression.Flowcytomctry was used to detect the cell cycle distribution.Results IC20 of LNCaP and PC-3 was 2.5 and 10.0 μmol/L,respectively.SER of panobinostat at IC20 was 1.37(D0 ratio)and 1.11(Dq ratio)for LNCaP cells,and 1.78(D0 ratio)and 1.17(Dq ratio)for PC-3 cells.Expression of γH2AX gradually decreased in the 2 Gy irradiation-alone cells standing for the DSB repair,while γH2AX expression was persistent in the combination group.Irradiation triggered a G2/M arrest 6-12 hours after irradiation in LNCaP and PC-3 cells.G2/M arrest was observed when cells were treated with panobinostat for 24 hours,however,no significant change concerning cell cycle distribution was showed when cells received further irradiation.Conclusions Panobinostat Call radiosensitize prostate cancer cells,which may be related with increased DNA DSB,inhibition of DSB repair and attenuation of cell cycle modulation after irradiation.

Ischemic cardiomyopathy (ICM) is due to coronary atherosclerosis cause long-term myocardial ischemia, make myocardial fibrosis diffuse, produce similar clinical syndrome like idiopathic dilated cardiomyopathy, which is one of a common clinical disease and causes increasing hazards to human health.Timely recovery of blood perfusion and coronary artery recanalization is one of the effective methods of treatment.However, it brings new damage while tissues and organs receive reperfusion, which is myocardial ischemia-reperfusion injury.In this paper, the current research progress in medicine myocardial ischemia-reperfusion injury and how to build a simple and efficient experimental animal models to mimic human myocardial ischemia-reperfusion injury were reviewed.

Ischemic cerebrovascular disease(ICVD) is temporary or permanent reduction of blood flow of the main arteries of the brain caused by cerebral transient,sudden,reversible neurological dysfunction.Timely recovery of blood supply to the brain is the most important way to save the patient's life,but at the same time gave restore blood perfusion brain tissue organs brought new damage,which is cerebeal ischemia reperfusion injury(CIRI).How to build an ideal,efficient,easy to operate brain ischemia-reperfusion injury in animal models to simulate the clinical study,and how to improve and innovate this model by researchers in recent years were reviewed in this article.

Objective To study the protective effect of the hypothermic brain-protection apparatus on hemorrhagic shock rabbits. Methods Applying the modified Wigger's shock animal model, we observed the effects of a self-developed brain-protecting apparatus on the survival time and vital signs of rabbits. Results Hypothermic brain protection could considerably reduce 60% lipid hyperoxide in brain tissues as compared with that in the control group. It could also decrease heart rate and respiration and hence reduce tissue oxygen consumption. Conclusion Hypothermic brain protection can attenuate the brain lesion and obviously prolong the survival time of hemorrhagic shock animals.

OBJECTIVETo investigate the changes in serum amylase (AMY) and lipase (LPS) and their clinical significance in patients with acute paraquat poisoning (APP).

METHODSThe clinical data of 62 APP. patients who were admitted to Guangzhou 12th People's Hospital in 2014 were retrospectively analyzed. With clinical death being regarded as the end point of observation, the 62 patients were divided into survival group (n=32) and death group (n=30). The fasting venous blood of the patients on the 1st, 3rd, and 7th day after admission were collected, and the AMY and LPS levels were determined; the obtained data were compared between the two groups and analyzed statistically.

RESULTSThe AMY and LPS levels of the survival group on the 1st, 3rd, and 7th day after admission were significantly lower than those of the death group (P<0.05); the AMY and LPS levels of the death group on the 3rd day were significantly higher than those on the 1st day after admission (P<0.05); the survival group demonstrated no significant changes in AMY and LPS levels (P>0.05).

CONCLUSIONChanges in AMY and LPS levels in APP patients are reliable indicators for the determination of poisoning severity and prognosis.

Amylases ; blood ; Humans ; Lipase ; blood ; Paraquat ; poisoning ; Poisoning ; blood ; mortality ; Prognosis ; Retrospective Studies

Objective To re-evaluate the prognostic value of the 6th edition of UICC/AJCC staging system in patients with nasopharyngeal carcinoma (NPC) treated with intensity-medulated radiation therapy (IMRT). Methods From February 2001 to March 2007, Clinical data of 570 NPC patients initially treated with IMRT in Cancer Center of Sun yat-sen University were reviewed and the long-term survival was analyzed according to T, N and overall stages. Results The median follow-up was 42 months. 184 patients were followed up to 5 years. The 5-year local recurrence-free survival (LRFS), distant metastasis-free survival (DMFS) and overall survival (OS) of the whole group were 93. 0%, 85.4% and 83. 3% ,respectively. No statistically significant difference of LRFS was detected between the either two of stage T_1, T_(2a) and T_(2b)(100%, 100% and 94. 5% ;T_1 vs. T_(2b), χ~2 = 1.92, P =0. 166 ;T_(2a) vs. T_(2b), χ~2= 0. 35, P =0. 555), stage T_(2b) and T_3 (94. 5% and 91.3% ;χ~2 = 2. 62, P = 0. 106), or stage T_3 and T_4 (91.3% and 89. 5% ; χ~2 = 1.55, P =0. 214). The 5-year DMFS of stage N_2 was similar with stage N_1 or stage N_3(80. 2%, 86. 2% and 61. 4% ; N_2 vs. N_1, χ~2=2.22, P=0.136;N_2 vs. N_3, χ~2= 1.92, P=0.165). No statistically significant difference of 5-year OS was observed among stage Ⅰ , Ⅱ_a and Ⅱ_b(91.7%, 100% and 95. 3% ; Ⅰ vs. Ⅱ_b χ~2 =0.32, P=0.574;Ⅱ_a vs. Ⅱ_b,χ~2-0.25, P=0.617), or between Ⅳ. And Ⅳ_b(67.9% and 75. 0% ;χ~2 = 0.25, P = 0. 616). Conclusions The 6th edition of UICC/AJCC staging system shows poor predictive value for the long-term survival of NPC patients treated with IMRT.

ObjectiveTo investigate the effects of partial hepatic ischemia/reperfusion (I/R) on choline acetyltransferase(ChAT) expression in hippocampal neuron in mice.MethodsOne hundred adult male mice,aged 2 months,weighing 20-25 g,were randomly divided into 5 groups (n =20 each): normal control group(group C),sham operation group(group S),groups I/R1,I/R2,I/R3.Partial hepatic ischemia was produced by clamping left hepatic artery and portal vein for 20,30,40 min respectively followed by reperfusion in groups I/R1,I/R2,I/R3.Passive avoidance task was performed with 10 mice in each group at 4-9 and 18-23 d after operation respectively.The animals were sacrificed and their brains were removed for determination of the expression of ChAT in CA3 of hippocampal neuron.ResultsCompared with group C,the latency was significantly shortened and number of errors increased in groups I/R1 and I/R2 at 4-7 d after operation and in group I/R3 at 4-9 d after operation,the expression of ChAT in hippocampal neuron down-regulated in groups I/R1,I/R2 and I/R3 at 9 d after operation( P ＜0.05 or 0.01).There was no significant difference in the latency and number of errors at 18-23 d after operation and the expression of ChAT in hippocampal neuron at 23 d after operation among groups C,I/R1,I/R2 and I/R3 ( P ＞ 0.05).Compared with group I/R1,the number of errors was significantly increased at 4 and 5 d after operation in group I/R2,the latency shortened at 4-6 d after operation and number of errors increased at 4-9 d after operation in group I/R3,and the expression of ChAT in hippocampal neuron down-regulated at 9 d after operation in groups I/R2 and I/R3 ( P ＜ 0.05 or 0.01 ).There was no significant difference in the latency and number of errors at 18-23 d after operation and the expresson of ChAT in hippocampal neuron at 23 d after operation among groups I/R1,I/R2 and I/R3 ( P ＞ 0.05).ConclusionPartial hepatic I/R can result in transient cognitive impairment in mice by down-regulating the expression of ChAT in hippocampal neuron.