AIM:To optimize the technical parameters indica-daisy dropping pill(Flos chrysanthemi indici) through controlling the influencing factors. METHODS:To take weight variation of pills,comprehensive quality and disintegration time limited as index,the above factors were observed by orthogonal test. RESULTS:Good technological parameters of indica-daisy dropping pill were as follows PEG6000∶PEG400=54∶6 as matrix,dimethicone as refrigerant.Temperature of drug fluids of 70 ℃,internal and external diameter of dropper within 6.2 mm and 9.0 mm,proportion of drug and matrix(1∶3),30 dropping per minute.The weight variation of pills was small,type quality was good and disintegration time limited was short.The water absorbability was smaller than that of granules. CONCLUSION:The finished products are of good quality and appropriate for mass production.

ObjectiveTo standardize the process in making the model of acute regional cerebral ischemia in Sprague-Dawley rats, and to economize time and material.MethodsRegional cerebral ischemia rat's models were induced and modified according to Koizumi's method.ResultsThe time duration was controllable and the volume of cerebral infarct was determined by adverting the approaches such as the preparation of suture, anaesthesia of the animal, and the details of surgical operation.ConclusionThe acute regional cerebral ischemic model in rats made by Koizumi's method is stable and reliable, and is easy for the beginner to carry out under limited conditions.

Pichia pastoris is a new eukaryotic expression system with great potentials, has unexampled advantages in expressing the heterologous proteins and has gotten more and more wide application. The following aspects were described in detail: the advantages of Pichia pastoris in the expression of heterologous genes, the mechanism of the heterologous gene integrations, the glycosylation of the expressed protein and the modification of the post-translated protein.

Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

Objective To investigate the effect of recombinant human erythropoietin(rh-EPO) on the nerve regeneration of adult rats sciatic nerves. Methods Tirty-six healthy male Wistar rats were involved and left sciatic nerve repaired model was used.The experimental rats were divided randomly into two groups:the EPO group and the control group,18 rats in each group.rh-EPO 3 000 U/kg was injected daily into the abdominal in EPO group,and normal saline was injected into the abdominal every day after operation in control group.On 4 and 8 weeks after operation,these items were determined,the sciatic function index (SFI),biomechanics examination,histological observation,electrophysiological examination,myelinated fibers density and sectional area measurement.Results On weeks 4 after operation,the SFI of EPO group and control group were-65.26 ± 3.42 and-70.83 ± 4.12,respectively,the maximum tensile resistance were (3.86 ± 0.29)N/mm2 and (3.38 ± 0.21 )N/mm2,the delayed ratio of latency of motor nerve were 2.34 ± 0.23 and 2.78 ± 0.29,and the recovery ratio of wave amplitude were 0.23 ± 0.05 and 0.14 ± 0.03 respectively.On eight weeks after operation,the SFI of EPO group and control group were-51.34 ± 2.98 and-57.23 ± 4.86,respcetively,the maximum tensile resistance were (4.67 ± 0.36) N/mm2 and (4.13 ± 0.32) N/mm2,the delayed ratio of latency of motor nerve were 1.32 ± 0.15 and 1.62 ± 0.21,the recovery ratio of wave amplitude were 0.41 ± 0.09 and 0.26 ± 0.07,the nerve fibers cross ratio were 0.57 ± 0.05 and 0.38 ± 0.03,and the recovery ratio of sectional area of myelinated fibers were 0.81 ± 0.06 and 0.58 ± 0.03,respectively.Those items in EPO group were significantly superior to those in the control group (P ＜ 0.05 ＝.Conclusion rh-EPO can promote the injured nerve regeneration and improve the recovery of their function.

Objective: To develop the Risk Assessment Scale of Severe Psychiatric Patients in Community and test its reliability and validity. Methods: A random sample of 860 severe psychiatric patients, which was selected from 8 communities in Chaoyang District of Beijing, completed the Risk Assessment Scale of Severe Psychiatric Patients in Community. The internal consistency reliability, the observer reliability, and the correlative coefficients between the total and items of the scale were analyzed, and the exploratory factor analysis was conducted. Results: (1) The Cronbach's coefficient of the scale was 0. 86, and the observer coefficient was 0.92. (2) The Spearman correlative coefficients between the total and items ranged from 0.40 ～ 0.56. (3) Exploratory factor analysis showed that the scale had 4 main factors, all of which could account for 68.14 percent of the whole variance, and the ten item loadings ranged from 0. 60 ～0.91. (4) The patient who scored higher than 35 was called high risk patient Causing trouble behavior of high risk patient was obviously higher than others. Conclusion: The results indicate that the Risk Assessment Scale of Severe Psychiatric Patients in Community has good reliability and validity. It can be used to assess the risk of severe psychiatric patients in community.

Objective To investigated the effects of IL -18 on middle ear allergic inflammation in rat model of OME .Methods Eighteen SD rats were randomly divided into three group :group A(control group ,n=12 ears) , group B (OME model group ,n=12 ears) ,group C (IL -18 injectiion group ,n=12 ears) .The rat model of OME was established by sensitizing with ovalbumin (OVA) and later challenging in tympanic bullae .Recombinant rat IL-18 (1 μg ,+0 .2 ml saline ,) were injected in group C at 1 ,2 ,7 ,8 ,15 ,16 day .At the same time sacle ,0 .2 ml sa-line ,instead of IL -18 ,were intraperitoneal injection in group A and B .The morphologic changes of the middle ear epithelial cells and inflammatory cells infiltration were observed under light microscope .The level of IFN -γand IL-4 in tympanic lavage fluid(TLF) were determined by ELISA .Results Pathological examination showed that middle ear mucosa inflammation and eosinophil infiltration in group C were no less severe than group B .The numbers of neutrophils in group C increased significantly compraring with group B (P<0 .05) .Numbers of eosinophils in group C were slightly increased comparing with group B (P>0 .05) ,while significantly greater than that in group A (P<0 .05) .ELISA showed that the level of IFN -γ in group C was stronger than that in group B and A (both P<0 .05) .As compared to the group A ,the expression of IL -4 in group B and group C were remarkably stronger (both P<0 .05) ,no significant difference was found between group B and group C (P>0 .05) .Addtionally ,there was no significant difference in the ratio of Th1 /Th2(IFN -γ/IL -4)between group B and group C (P>0 .05) . Conclusion IL -18 acts as an immune regulatory factors ,significantly increases Th1 cytokine IFN -γ.Although to some extent alleviate the OME rat middle ear Th1 /Th2 imbalance ,there is still excessive activation of Th cells . Th1 and Th2 cells factor are excessive for the secretion disorder of the immune response status .The OME rat mid-dle ear allergic inflammation has not been fundamentally alleviated ,the underlying mechamism should be further studied .

Bacterial N-Acyl-L-homoserine lactones (AHLs)-mediated quorum sensing is involved in the regulation of diverse biological functions. As the bacterial pathogen population density increases, AHLs concentration secreted by pathogen reaches a threshold and then interacts with their intercellular receptor and triggers expression of virulence genes. It is a promising approach to biologically control bacterial disease in plants and animals by manipulating bacterial AHL-quorum sensing with AHLs-degrading enzyme and AHL analogue.

Relapse, which puzzled several generations of hematologists, is the bottle-neck of radical treatment for leukemias. The progress of Human Microbiome Project at the beginning of 21st century suggested that human body was a super-organism constituted by the core of human cells and symbiotic microorganisms. The elucidation and characterization of endogenous retrovirus and prion protein suggested the possible effects of co-evolutional microorganisms on human health. Recently, the elucidation of the roles of tunneling nanotubes in intercellular communication and transportation suggested a novel way for cellular communication and transport of oncogenic materials. The role and significance of in vivo cell fusion have been studied in more detail. On the other hand, donor cell leukemia was reported. All of these approaches provide novel insights for studying the mechanism of leukemia relapse. Based on previous work, the authors suggest the hypothesis: there are two possible mechanisms for the relapse of leukemias: the minimal residual disease (MRD) and intercellular transportation of oncogenic materials.
Cell Fusion ; Humans ; Leukemia ; pathology ; Neoplasm, Residual ; pathology ; Recurrence

OBJECTIVETo study the effect of transforming growth factor-β activated kinase-1 (TAK1) gene silencing on the proliferation and apoptosis of Kasumi-1 cells induced by arsenic trioxide (As₂O₃).

METHODSAcute myeloid leukemia with t(8;21) cell line Kasumi-1 cells were treated with As₂O₃ or in combination with TAK1 siRNA interference technology. The experiment was divided into four groups: Kasumi-1 cells without any treatment, TAK1 specific siRNA transfection alone, Kasumi-1 cells treated with different concentration of As₂O₃, TAK1siRNA transfection combined with As₂O₃. CCK-8 was used to detect the cell viability. The expression of phosphorylated c-Jun N-terminal kinase (P-JNK) was determined by Western Blot. Cell apoptosis and growth were examined by morphological and colony formation assay.

RESULTSAfter Kasumi-1 cells were treated with As₂O₃, the rate of cell inhibition was concentration-dependent, and the 50% inhibitory concentration was 3.5 μmol/L. The highest expression level of P-JNK appeared in 30 minutes after cells were treated with As₂O₃. The apoptosis rates of Kasumi-1 cells without any treatment, TAK1 siRNA interference alone group, As₂O₃ alone group and the combined group were (5.02 ± 1.13)%, (6.18 ± 0.28)%, (48.33 ± 2.70)% and (86.07 ± 2.21)%; colony formation rates were (73.83 ± 2.78)%, (76.03 ± 1.46)%, (55.07 ± 1.50)% and (22.20 ± 1.15)%; apoptosis rate of TAK1 siRNA group and the untreated group has no significant difference (P = 0.052); colony formation rate between TAk1 siRNA group and the untreated group has no significant difference (P = 0.179), but the difference in other groups was significant (P = 0.000).

CONCLUSIONSilencing the expression of TAK1 can enhance the anti-proliferative and pro-apoptotic effect of As₂O₃ on Kasumi-1 cells, and its mechanism may be through the TAK1 downstream JNK signal pathway.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Leukemia, Myeloid, Acute ; enzymology ; pathology ; MAP Kinase Kinase Kinases ; genetics ; metabolism ; Oxides ; pharmacology ; RNA Interference ; RNA, Small Interfering ; genetics ; Signal Transduction