OBJECTIVE:To prepare diphenhydramine hydrochloride gel and establish its quality control.METHODS:The best formula was optimized by orthogonal design with sodium carboxymethyl cellulose and so on as the base materials,the content of diphenhydramine hydrochloride was determined by UV-spectrophotometry at the wavelength of258nm.RE?SULTS:The diphenhydramine hydrochloride gel was even,fine and which has a good dispersivity;In the concentration range of100～400?g/ml,the diphenhydramine hydrochloride had a linear relationship(r=0.9999,n=6),the average recovery was99.95%,RSD=0.95%(n=9).CONCLUSION:The preparation was stable in quality and reliable in quality control method.

OBJECTIVE:To optimize the preparation technique of levofoxacin lactate and glucose injection.METHODS:Orthogonal test was applied in which orthogonal table L 16 (4 5 )was chosen to perform the experiment.The effects of4factors,i.e.the pH value,amount of activated carbon added,temperature of heating and time of heating on the content of levofloxacin lactate was investigated.RESULTS:The optimum preparation technical conditions were turned out to be the following:the pH value was4.2～4.8,the amount of activated carbon was0.1%,the heating temperature was50℃with10min of heating time.CONCLUSION:The quality of the finished products from this preparation technique was in line with quality requirement.

OBJECTIVE:To prepare lomefloxacin hydrochloride and diethylstilbestrol oil and to establish the method for its quality control.METHODS:The content of lomefloxacin was determined by UV-spectrophotometry at the wavelength of283nm.RESULTS:The detected concentration of lomefloxacin hydrochloride was in linear relation to absorptivity in the range of2.5～16.0?g/ml(r=0.9995,n=6).The average recovery was99.8%(RSD=0.74%).CONCLUSION:The preparation technique is simple and the method for quality control is feasible.

Objective:To explore the possible specific perinatal risk factors in the development of autism through comparing the perinatal risk factors between patients with autism and other psychotic disorders.Methods: In this retrospective research,197 cases with autism and 93 cases with other psychotic disorders were selected and assessed with the self-made scale of perinatal risk factors.Statistical analyses were performed using t test and Chi-square test.Results:Compared with the control group,the autistic subjects had a significantly higher frequency of the perinatal risk factors(51 % vs.68%,P=0.003),particularly the rate of catching a cold during their mothers' pregnancy(14.2% vs.6.5%,P=0.038)and the rate of prematurity(10.7% vs.3.2%,P=0.022)than the control subjects.Feeding methods was different between the two groups(P=0.038).Conclusion:Catching a cold during mothers' pregnancy,prematurity and feeding methods may be related to the development of autism.

Objective To develop a new method for the determination of trace silver in water by flame atomic absorption(FAAS) after cloud point extraction.Methods Diethyldithiophosphate(DDTP) and triton X-114 were respectively used as the chelating agent and surfactant.The effects of experimental conditions such as acidity,concentration of chelating agent,surfactant and methanol,equilibration temperature and time,interference ion on cloud point extraction were investigated.Results Under the optimum condition,0.5 ml of 0.1 mol/L DDTP solution,0.5 ml of 50 g/L Triton X-114 and 5 ml of 1 mol/L hydrochloric acid were added and the volume was made up to 50 ml.The mixture was heated in a thermostatic bath at 40 ℃ for 15 min,the linear range of determination for silver was 0-100 ng/ml,the equation of linear regression was A = 0.005 8cAg+ 0.001 6,r = 0.999.The detection limit was 0.83 ng/ml,the recovery rate was in the range of 97%-105%,and the relative standard deviation was 3.1%(n=11) for 20 ng/ml Ag.There was no significant interference for silver solution containing 5 000 times of K+,Na+,Ca2,Mg2,NO3,SO4 + +-2-and Cl-,1 000 times of Al3+ and Zn2+,500 times of Fe2+ and Mn2+,100 times of Pb2+ and Cu2+.Conclusion The method is rapid,accurate,simple and applicable to the determination of trace silver in water samples.

Taking Coxsackie B3 virus cDNA labelled by Biotin as detecting probes and 25 kinds of standard enteroviruses as detected viusr, a spot nucleic acid hybridization method has been established to detect enterovirus RNA gene. The method has simple operation and its sensitivity is 50-80 TCID50 spot. Reagent is safe and stable, and may be stored for long period. pCBHⅢ/35.51 probe used has wider hibridization spectrum to enteroviruses; pCBⅢ/29 only reacts with RNA of Coxsackie B3 virus in strict condition. Consequently, the hybridization character which these probes possess is suitable to determining enlerovirus in clinical samples.

Objective: To investigate the expressions of programmed death ligand 1（PD-L1）in triple-negative breast cancer (TNBC) and its correlation with angiogenesis. Methods: 120 cases of TNBC patients who underwent surgery in the Fourth Hospital of Hebei Medical University from March 1, 2011 to June 1, 2012 were collected. The tumor tissues of patients were surgically resected and confirmed by pathology. PD-L1 protein expression in TNBC tissues of 120 patients was detected by tissue microarray combined with immunohistochemistry, and its relationship with various clinical indicators was analyzed. Blood vessels and lymphatic vessels were labeled withCD34andD2-40todetectmicrovesseldensity(MVD)andlymphaticvesseldensity(LVD)inTNBC.Results:Thepositiveexpression rate of PD-L1 in the tumor cells and interstitial infiltrating lymphocytes fromTNBC was 56.7% (68/120); No correlation was found between PD-L1 protein expression and the gender, age, histological grade, clinical stage, or tumor size of patients with TNBC (P>0.05), but related to the lymph node metastasis (P<0.05) and vascular thrombus (P<0.05). TNBC with high PD-L1 expression exhibited high incidence of lymph node metastasis and formation of vascular thrombus, and the expression of PD-L1 was positively correlated with MVD (r=0.500, P=0.02) as well as LVD (r=0.662, P=0.01). Log-Rank test showed that the survival time of TNBC patients with positive PD-L1 protein expression was significantly shorter than that of patients with negative expression (P<0.05). Cox multivariate analysis suggested that PD-L1 protein expression could be an independent prognostic factor for TNBC overall survival. Conclusion: PD-L1 plays an important role in TNBC angiogenesis and lymphangiogenesis, and is closely related to TNBC invasion and metastasis; blocking PD1/PD-L1 signal pathway is expected to be an effective new strategy for TNBC treatment.

Objective To study the Janus Kinase 2 V617F (JAK2V617F) point mutation in bcr-abl-negative myeloproliferative disorders (MPD) and explore its clinical significances. Methods Genomic DNA was isolated from bone marrow or peripheral-blood granulocytes. Allelespecific-polymerase chain reactions (AS-PCR), restriction enzyme digestion in combination with PCR product sequencing were performed to detect the mutation in genomic DNA. 110 patients were detected, including 41 with bcr-ablnegative MPD, 25 with bcr-abl-positive chronic myelogenous leukemia (CML), and 44 with acute leukemia.Results JAK2V617F was presented in 11 cases(91.7 %) of 12 polycythemia vera (PV), 8 cases(53.3 %) of 15 essential thrombocythemia(ET), 4 cases (57.1%) of 7 idiopathic myelofibrosis (IMF), while in other patients including 7 hypereosinophilic syndrome (HES), 25 bcr-abl-positive CML, 24 acute myelocytic leukemia (AML), 18 acute lymphoblastic leukemia(ALL), and 2 acute mixed lineage leukemia, JAK2V617F can not be detected. All positive samples and 10 negative samples identified by AS-PCR and restriction enzyme digestion were confirmed further by DNA sequencing. Conclusion The frequency of JAK2V617F mutation was more than 90 % among patients with PV, more than 50 % among patients with ET and IMF. The detection of JAK2V617F mutation will be of great significanees in the diagnosis and differential diagnosis of MPD. This mutation can be a molecular marker of MPD and might be a treatment target in the future.

Objective:This work aims to detect the levels of miR-200c/141 methylation and miR-200c/141 in gastric cancer tissue and investigate the relationship between miR-200c/141 expression and clinical parameters. Methods:The methylation status of miR-200c/141 CpG island and miR-200c/141 in gastric cancer tissue specimens was evaluated by qRT-PCR or BS-MSP method. We analyzed the relationship among the methylation status of miR-200c/141 CpG island, expression level of miR-200c or miR-141, and clinical parame-ters. Results:The status of miR-200c/141 CpG island methylation in gastric cancer tissue was significantly higher compared with that in paracarcinoma tissue. MiR-200c and miR-141 were markedly decreased in gastric cancer tissue compared with those in adjacent tis-sue. MiR-200c/141 CpG island methylation was negatively related with the expression of miR-200c and miR-141 in gastric cancer speci-mens. Conclusion:The upregulation of miR-200c/141 CpG methylation inhibits miR-200c/141 expression in gastric cancer tissue.

Objective To evaluate the effect of sodium nitroprusside(SNP)and N-nitro-l-arginine-mythel-ester (L-NAME) on apoptosis of spermatogenic cells in rats. Methods Fourty adult male Sprague-Dawley rats (60-70days) were divided into four groups.Each group was injected intraperitoneally with one of the following agents, once a day, for 12 days: 1. SNP; 2.L-NAME;3.SNL+L-NAME;4.Normal saline NS group.Two hours after the last time injection the rats were sacrificed.TUNEL staining and flow cytometry analysis were used to detect the apoptosis of spermatogenic cells. Results Sub-monoploid and apoptosis index (AI) in SNP group was significantly higher than that of NS group and sub-monoploid and apoptosis index (AI) in L-NAME group were significantly lower than that of NS group by FCM and TUNEL (P0.5) was found.Conclusion SNP can accelerate the apoptosis of spermatogenic cells and L-NAME can inhibite the apoptosis of spermatogenic cells,The effect of SNP and L-NAME on apoptosis of spermatogenic cells probably occurs through the action of nitric oxide.