In order to explore the significance of cardiomyocyte apoptosis in the early myocardial ischemia,the rat model of acute myocardial ischemia was established,and the apoptotic cells were detected in the early phase of ischemia(within 6h)with TUNEL method.The results showed that scanty apoptotic cells could be observed in the ischemic region 1h after ischemia,and reached the peak 3h after ischemia,then decreased.No apoptotic cells were found in the normal region.In the peri ischemic region,apoptotic cells could also be observed 1h after ischemia,and reached the peak 5h after ischemia.It is indicated that apoptosis is the major form of early ischemic myocardial damage,and the detection of apoptotic cardiomyocyte may provide a new sensitive and objective method for the postmortem diagnosis of early myocardial infarction.

To study the expression of both Fos and HSP70 proteins in the early stage of acute myocardial ischemia/reperfusion(I/R) in rats with immunohistochemistry in order to search the objective morphologic evidence for the forensic pathological diagnosis.I/R rat model was established by ligation of the anterior branch of the left coronary artery.All animals were divided into 7 groups.Tissues were cut and stained immunohistochemistry with the corresponding anti Fos and anti HSP70 sera as the first antibodies.There was no expression of both Fos and HSP70 proteins in control group.The expression of HSP70 protein began to increase in myocytes 1h after I/R in the ischemic areas and increased gradually wtih the prolongation of ischemia,while it began to express 2h after I/R in the non ischemic areas.Fos protein began to express 0 5h after I/R in the ischemic areas,1h in the non ischemic areas.The expressions of both HSP70 and Fos proteins in the non ischemic areas were weaker than those in the ischemic areas.Meanwhile,the expression of HSP70 and Fos proteins appears firstly in the inner layer of myocardium and extended gradually toward the outer layer.The expression of Fos protein mainly located in the inner layer of myocardium 0 5h after I/R and then extened to the whole layer of myocardium 1h after I/R.The expression of Fos protein in outer layer was stronger than that in inner layer at 4h after I/R.The expression of HSP70 protein located mainly in the inner layer of the myocardium at 1h and 2h after I/R,and then extended to the whole layer of the myocardium at 4h and 6h after I/R.The results indicated that the I/R induced the expression of both HSP70 and Fos proteins in the early stages of myocardial ischemia.The location and the intersity of the immunoreactivity of these two proteins changed at the different stages after I/R.These changes observed in the present study might be useful for the forensic pathological diagnosis of the early myocardial ischemia.

Objective To analysis the fresh motherwort capsule treatment of postpartum uterine hemorrhage clinical value.Methods100 cases of postpartum uterine hemorrhage patients of our hospital from January 2016 to December 2016 were selected as the research object, the control treatment, use contractions observed group of using fresh motherwort capsule combined contractions for treatment.Check after drug treatment in patients with uterine instauration and clinical curative effect.ResultsAfter five days, two groups of patients with postpartum uterine instauration is close, there was no statistically significant difference.After 14 days, observation group of patients with uterine instauration was significantly better than the control group, there was statistically significant difference (P< 0.05);Observation group of patients treatment of cure rate (76.0%), and total effective rate (96.0%) were significantly higher than control group (56.0%, 88.0%), statistically significant difference (P< 0.05).ConclusionFresh motherwort capsule in the treatment of postpartum uterine hemorrhage has significant clinical efficacy, can promote the patients with uterine instauration, has significant clinical value, worthy of popularization and application.

Objective To investigate the potential of substrate elasticity in regulating rapid differentiation of HepaRG cells into hepatocyte-like cells ,and further provide hepatocytes for bioartificial liver. Methods The substrate elasticity was divided into 4 groups. The expressions of albumin(ALB)were detected by albumin-green fluorescent protein-reporter system (ALB-GFP-reporter system) and Image J software;the cell morphology was observed by microscope and the amounts of cell were detected by cell Titer-Blue cell viability assay kit (alamar blue). Results The results of ALB showed that at the 4th hour,the expressions of ALB inside the HepaRG cells between 4s group and 8s group,16 s group and Glass group were not statistically different (t = 0.791,1.389, 2.481,P>0.05);at the 4th day,the expressions of 4s group had statistical differences in comparison with those of 16s group and Glass group(t = 12.41,12.52,P < 0.05),but not statistical difference when compared with those of 8s group(t = 2.603,P > 0.05);at the 7th day,the expressions of 4s group were statistically different from those of 8s group,16s group and Glass group(t=3.266,6.725,8.005,P<0.05). The microscope showed that only a typical epithelial morphology was observed before differentiation ,then hepatocyte-like cells and bile duct-like cell were observed at differentiated phases. The alamar blue showed that at the 4th and the 7th day,the cell number of 4s group were not statistically different from that of 8s group,16s group and Glass group(P >0.05). Conclusion Soft substrate can promote differentiation of HepaRG cells.

Aim To explore the toxicity and safety of five kinds of known positive drugs, cyclophosphamide, acetyl salicylic acid, tetracycline hydrochloride, dexa-methasone acetate and azacitidine, using zebrafish em-bryos. Methods We selected normally developed 4 hpf zygote, and used water bath infecting method to add the drug to the artificial seawater. Each drug had five concentrating groups, a separate control group and solvent control group. We observed the dead zebrafish embryos after 120 hpf drugs, counted the number of deaths and deformities of zebrafish embryos, and cal-culated mortality abnormal rate, the median lethal con-centration (LC50 ), concentration for 50% of maximal effect (EC50 ), therapeutic index (TI) under 120 hpf condition. We also used the formula TI = LC50 / EC50 to calculate positive drug therapeutic index. Based on measured LC50 we calculated most nonlethal concentra-tion (MNLC) of each drug setting, namely 1 / 10 MN-LC, 1 / 3 MNLC, MNLC,LC10 four concentration, tha-lidomide as a positive control, vitamin C as a negative control, artificial seawater as control, 0. 5% DMSO as solvent control. Put in 28. 5 ℃ environment for 120 hours,embryo development was observed daily for de-velopmental state,mortality,deforming rate and abnor-mal condition. Results The result of five drugs LC50 in descending order: cyclophosphamide > azacitidine> tetracycline hydrochloride > acetylsalicylic acid >dexamethasone acetate. EC50 in descending order: cy-clophosphamide > tetracycline hydrochloride > azaciti-dine > acetylsalicylic acid > dexamethasone acetate. The TI values of cyclophosphamide, acetyl salicylic acid, tetracycline hydrochloride, dexamethasone ace-tate, azacitidine were 1. 92, 1. 11, 1. 05, 1. 44, 2. 99, respectively. Conclusion Zebrafish embryo model can be used in the preliminary evaluation of drugs, and the study of early developmental toxicity and safety.

Objective To evaluate the efficacy and safety of Sheng-qi-zhuang-yang formula combined with specific immunotherapy (SIT) with standardized house dust mite vaccine on allergic asthmatic children. Methods Participants were 100 children with mild to moderate allergic asthma , who were receiving SIT at Guangdong Provincial Hospital of Chinese Medicine from Jan 2011 to Dec 2013 , who were divided into treatment group and control group. Patients in the treatment group received Sheng-qi-zhuang-yang Decoction combined with SIT while patients in the control group received SIT alone. Asthma symptom scores, respiratory function and related adverse events were compared before and after treatment, between groups. Results The desensitization treatment functions was ahead of time than expected in both groups. There is no significant difference between groups in terms of respiratory function and adverse effects. Conclusion Sheng-qi-zhuang-yang decoction combined with SIT for allergic asthmatic children seems to advance clinical effect without increasing adverse events. Further large scale clinical trial is required to confirm this.

Objective To analyze the clinical effects of thrombolytic therapy in patients with ischemic in-hos-pital stroke (IHS). Methods The clinical data were collected from patients with ischemic IHS in the last five years. The patients were divided into thrombolysis group and non-thrombolysis group, according to the use of recombinant tissue plasminogen activator (r-tPA) treatment. The clinical outcomes were measured by the modified Rankin scale (mRS) at discharge. Results There were a total of 121 patients in this study. There were 6 patients in thrombolysis group and 115 patients in the non-thrombolysis group, respectively. Six patients (100%) in the thrombolysis group achieved favor-able outcomes (mRS 0~2) at discharge whereas only 42 patients (36.5%) in the non-thrombolysis group achieved fa-vourable outcomes. The rate of favorable outcomes was significantly higher in the thrombolysis group than in the non-thrombolysis group (P<0.05). Conclusions R-tPA thrombolytic therapy can improve the prognosis of patients with ischemic IHS.

Objective To investigate radiation risk perception and nuclear acceptance after the 60Co stuck source incident in June 2009 in Qixian County,Henan Province.Methods From March to April 2010,23 hamlets and 2 schools from 5 towns in Qixian County were selected,while 3 hamlets and 2 schools from 3 towns were used as control in Huixian County,1 35 km away where no any impact of the incident was found.Face-to-face questionnaire survey was conducted among three groups including mass public,elite public(representatives of public opinion,such as physicians,teachers,govemmental officials,administrative personnel,private entrepreneurs),and middie school students(the third grade students in junior middle schools and the first grade students in senior middle schools).Results A total of 1 340 valid questionnaires were collected.The public knowledge levels about radiation and nuclear power were low among all groups in two areas and public knowledge levels in Qixian County were higher than those in Huixian County.In both areas.the knowledge levels of the students were higher tIIan those among the elite public,and that among the general public were the lowest.The knowledge levels of males and those with higher educational levels were higher than those of females and those with lower educational levels.More than 40.0% of the respondents supported the idea to build nuclear power plants in China.but only a small part of them supported to build nuclear power plants in their own areas.The percentages of opponents against local nuclear power plant construction were especially higher among the students and those with higher educational levels.More than 60% of the mass public and elite public believed the explanation of the government.More than 80% of the respondents held negative attitudes towards the performance of the Qixian County Government in dealing with this incident.The student gave the lowest evaluation of the credibility and the specific performance of the local government,whereas the elite public placed themselves between the students and the mass public.The reasons why the local people left their homes to evade risk included rumors(67.2%),lack of radiation knowledge(61.9%),group psychology(59.2%),and failure of the government to inform the truth to the public in time(42.3%).Conclusions The public still fails to know enough about radiation and nuclear power.Although they trust the government,they are dissatisfied with the performance of the local government in dealing with the incident.The negative evaluation of the students is worth concerning.

Pyrosequencing is a tool based on bioluminescence reaction for real-time analyzing DNA sequences. The sensitivity of pyrosequencing mainly depends on luciferase in reaction mixture. However, the instability of pyrosequencing reagents caused by fragile wild Photinus pyralis luciferase (PpL) in conventional pyrosequencing usually leads to unsatisfied results, which limits the application of pyrosequencing. In order to improve the stability of pyrosequencing reagents, the coding sequences of mutant thermostable Luciola lateralis luciferase (rt-LlL) was synthesized, and inserted into the plasmid of pET28a(+) to express the thermostable rt-LlL with a 6 x His-tag in the N terminal. The purified rt-LlL with the molecular mass of 60 kDa was obtained by Ni-affinity chromatography. The specific activity of rt-LlL was determined as 4.29 x 10(10) RLU/mg. Moreover, the thermostability of rt-LlL was investigated, and the results showed that rt-LlL had activity at 50 degrees C, and remained 90% of activity after incubated at 40 degrees C for 25 min. Finally, rt-LlL was used to substitute commercial Photinus pyralis luciferase in conventional pyrosequencing reagent to get thermostable pyrosequencing reagent. Comparing with conventional pyrosequencing reagent, the thermostable pyrosequencing reagent is more stable, and it's activity would not lose when incubated at 37 degrees C for 1 h. This study laid foundation of establishing reliable and stable pyrosequencing system which would be applied in Point-of-Care Testing.
Animals ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Fireflies ; enzymology ; Luciferases ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; methods

We expressed recombinant single-stranded DNA-binding protein (r-SSBP) from Escherichia coli with the molecular weight of 24-kDa by using genetic engineering strategy, and demonstrated the single-stranded DNA (ssDNA)-binding activity of r-SSBP by electrophoretic mobility shift assay (EMSA). To further characterize r-SSBP, we studied the effects of r-SSBP on melting temperature (T(m)) of DNA. The results showed that r-SSBP could bind to ssDNA, and lower the T(m) of DNA, especially for single-base mismatched DNA. Therefore, r-SSBP significantly increased the T(m) difference between single-base mismatched DNA and perfect matched DNA. These results are very beneficial for single-nucleotide polymorphism detection. Moreover, we applied r-SSBP in high sensitive pyrosequencing system developed by our group. The results suggest that the r-SSBP decreased non-specific signals, corrected the proportion of signal peak height and improved the performance of pyrosequencing.
DNA-Binding Proteins ; biosynthesis ; genetics ; Diphosphates ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; methods