Objective To investigate the recurrent etiologic causes in otitis media with effusion(OME).Meth-ods The patients with OME had been conducted clinical evaluation and ventilation tube insertion.Among 255 cases observed,85 cases were recurrence within one year,while 170 cases were recovered.Factors including age,gender, course of disease,smoking environment,recurrent respiratory tract infection,chronic nasal sinusitis,nasal struc-tural abnormalities,adenoid hypertrophy/nasopharyneal lymphadenosis,and mastoid gasification condition,histo-ry of tube insertion,duration of ventilation tube were evaluated by univariate analysis and multivariate logistic re-gression analysis and compared among preschool children,school age children and adults.Results In univariate a-nalysis,it was strongly associated with age,higher prevalence of recurrent respiratory tract infection(χ2= 22.546,P= 0.000),chronic nasal sinusitis(χ2= 5.211,P= 0.023),adenoid hypertrophy/nasopharyneal lymphadenosis(χ2= 10.338,P= 0.002),mastoid pneumatization adverse(χ2= 15.196,P= 0.000)and duration of ventilation tubeand (χ2= 11.347,P= 0.001).In multivariate logistic regression model,five of these factors were found to be predictors of recurrent OME,which were younger age,recurrent respiratory tract infection (P= 0.001,OR= 2.992),adenoid hypertrophy/nasopharyneal lymphadenosis (P= 0.021,OR= 2.198),mastoid pneumatization adverse (P= 0.000, OR= 3.433)and duration of ventilation tube(3~6 m)(P= 0.010,OR= 2.237).When comparing the difference a-mong preschool children,school age children and adults,recurrent respiratory tract infections in preschool and school age children had the statistic significance(P<0.05),and adenoid hypertrophy in preschool had the statistic significance(P<0.05).Conclusion Younger age,recurrent respiratory tract infection,chronic nasal sinusitis,ade-noid hypertrophy/nasopharyneal lymphadenosis,mastoid pneumatization adverse and less time of ventilation tube are the risk factors causing recurrence of OME.When comparing the recurrent preschool children,school age chil-dren and adults cases,more recurrent respiratory tract infection in preschool and school age children have statistic significance,and adenoid hypertrophy in preschool has the statistic significance.

Objective To study clinical characteristics and treatment effects on old patients with sudden sen-sorinerural hearing loss .Methods To compare clinical characteristics and effects of 357 cases of unilateral SSHL from Sep ,2013 to Mar ,2015 in the First Affiliated Hospital of Zhengzhou University .The patients were divided into 2 groups :the old group(66 cases)≥60 years ,and the control group(291 cases)<60 years .All cases were trea-ted with drugs to supply nutrition to the nerves and drugs to improve circulation and intratympanic injection of dexa-methasone .Results The genders ,ear sides ,concomitant symptoms and onset time ,degrees of hearing in 2 groups showed no significant difference (P>0 .05) ,but there were less patients with the type in low frequencies (1 .51% ) and more with the type in high frequencies (22 .73% ) and the type in total deafness (50 .0% ) than those of in the control group(14 .43% ,15 .44% ,39 .52% ) .The old group had more patients with systemic diseases (60 .61% ) and the hearing impairment of the other ear (57 .58% ,P<0 .05) .Total effective rate was 70 .03% ,which the old group and the control group were 60 .61% and 72 .16% ,respectively .The difference had no statistical significance (P>0 .05) .The old group's complete recovery rate was 10 .61% ,which was significantly lower than that in the control (P<0 .05) .Conclusion The old group has more patients with systemic diseases and the hearing impairment of the other ear .Their hearing impairment more easily occurs in the type in high frequencies and the type in total deafness . But the complete recovery rate is low ,the doctors and patients should have a reasonable expectation .

Objective:To research the number and function of monocyte-macrophages in patients with chronic active hepatitis B. Methods:The 51 chronic viral hepatitis B( CHB) patients were selected randomly,which consisted of 20 cases of mild-moderate,31 cases of severe group and 13 cases of healthy controls. PBMCs were separated by percoll. Monocytes were tagged by CD14,the molecules CD80,CD86,HLA-DR and CD163 were detected by flow cytometry which expressed on the surface of PBMCs. Serum cytokine were detected for IL-10, IL-12 and IL-23 by ELISA. The distribution of CD68 was detected in the liver by immunohistochemical staining. Results:The expressions of CD80 for all chronic hepatitis B patients were lower than the controls respectively,no matter mild-moderate or even severe group. Similarly,the HBV patients expressed lower level of CD86 in the peripheral blood mononuclear cells when compared with the control group. Furthermore, there was statistically difference between the levels of CD86 in severe group compared with control group (P<0. 01). As the expression of CD80 and CD86,the levels of HLA-DR in the patents had also declined when compared with controls. While the HLA-DR levels in both the mild-moderate HBV hepatitis groups were statistically significant higher than the severe group (P<0. 01). Different from the above all,the expression of CD163 in all chronic HBV hepatitis was higher than the control group. The CD68 positive cells in chronic HBV patients were observed and infiltrated increasingly in portal area and hepatic lobules (P<0. 05). There were statistically significant differences of IL-10 levels between the mild-moderate group,severe group and the control group,respectively (P<0. 01). Conclusion:Macrophages have participated in the pathological lesions of liver in CHB patients,among peripheral blood mononuclear cells,the phenomena of imbalance between type M1/M2 and polarization to type M2 have been observed,which participated in the development of the chronicity of CHB.

Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.

Objective To investigate the relationship between dyslipidemia,obesity,insulin resistance (IR)and various degrees of non.alcoholic fatty liver disease(NAFLD)in patients with type 2 diabetes mellitus (T2DM), and the risk factors of NAFLD.Methods Two hundred and sixty-eight patients were divided into three groups(non-NAFLD group,mild NAFLD group,moderate and severe NAFLD group)by liver ultrasonography.Body height(H),weight(W),waist circumference(WC),hip circumference(H)were measured.The levels of fasting blood glucose (FBG),glycosylated hemoglobin A_1c(GHbA_1C),serum total cholesterol(TC),serunl high density lipoprotein(HDL-C),serum low density lipoprotein(LDL-C),serum triglyceride (TG),alanine aminotransferase (ALT)and fasting serum insulin(FINS)were measured.Body mass index(BMI),the waist to hip ratio(WHR)and insulin resistance index(HOMA-IR)were calculated.Unconditional logistic regression model was used to test for the risk factors of NAFLD.Results BMI、WC、WHR、HNS、HOMA.IR、TC、LDL-C、TG and ALT in NAFLD group were significantly higher than those in non-NAFLD group (P<0.05).The levels of BMI、WC、WHR、HNS、HOMA-IR、 TG and ALT increased significantly in moderate and severe NAFLD group compared with mild NAFLD group(P<0.05).TG、WHR and HOMA.IR were the risk factors of NAFLD(P<0.05,OR=2.394,3.273,5.256).Conclusions NAFLD in patients with T2DM had remarkable dyslipidemia,overweight,central obesity and insulin resistance.TG、WHR and HOMA.IR were risk factors of NAFLD.

[Objective] To investigate the clinical characteristics and risk factors of lower-extremity arterial disease in the patients with newly diagnosed type 2 diabetes mellitus combined with nonalcoholic fatty liver disease (NAFLD). [Methods] One hundred fifty-one patients were investigated respectively. The patients were divided into two groups (NAFLD-Group and non-NAFLD group) by liver ultrasonography and disease history, then their clinical data were collected and compared in order to find the differences of biochemical indicators and the morbidity of lower-extremity arterial disease between two groups. [Results] Ninety-two cases (60.93%) were complicated with NAFLD. NAFLD group had higher levels of fast insulin and C peptide level, postprandial insulin and C peptide level, uric acid, body mass index (BMI), homeostasis model assessment (HOMA-IR) and lower level of high-density lipoprotein cholesterol and insulin sensitive index than those of without NAFLD (P<0.05). One hundred and one cases(66.89%) were complicated with lower-extremity arterial disease. The morbidity of lower-extremity arterial diseases was higher in NAFLD group than that of without NAFLD group (75% vs. 54.24%, P<0.01). [Conclusion] Both lower-extremity arterial disease and NAFLD are common complicated with type 2 diabetes. The morbidity of lower-extremity arterial diseases was higher in NAFLD group than that of without NAFLD group.

Soil water is one of the most important components in hydrological cycle. The stable hydrogen and oxygen isotopes in soil water have been increasingly used in the ecological, environment and hydrological research. In view of different techniques for extracting soil water, there is significant difference in theδD andδ18 O composition. This paper presents a method for analyzing hydrogen and oxygen isotopes in soil water by using elemental analyzer and isotope ratio mass spectrometry with accelerated solvent extraction for sample pretreatment. The conditions are: extraction solvent: dichloromethane, temperature: 100 ℃, pressure of 10. 3 MPa, static time:10 min. The samples were extracted three times, and with cycle values of four, four and three, respectively. Comparing with the added water, the deuterium and oxygen isotope values in the extracted soil water enrich 2. 12‰-4. 58‰ and 0. 17‰-0. 93‰, respectively. The reproducibility of replicate extractions of soil water is around ±0. 89‰ for δD and ±0. 37‰ for δ18 O.

A novel targeting drug carrier (FA-BO-PAMAM) based on the PAMAM G5 dendrimer modified with borneol (BO) and folic acid (FA) molecules on the periphery and doxorubicin (DOX) loaded in the interior was designed and prepared to achieve the purposes of enhancing the blood-brain barrier (BBB) transportation and improving the drug accumulation in the glioma cells. 1H NMR was used to confirm the synthesis of FA-BO-PAMAM; its morphology and mean size were analyzed by dynamic light scattering (DLS) and transmission electron microscope (TEM). Based on the HBMEC and C6 cells, cytotoxicity assay, transport across the BBB, cellular uptake and anti-tumor activity in vitro were investigated to evaluate the properties of nanocarriers in vitro. The results showed that the nanocarrier of FA-BO-PAMAM was successfully synthesized, which was spherical in morphology with the average size of (22.28 ± 0.42) nm, and zeta potential of (7.6 ± 0.89) mV. Cytotoxicity and transport across the BBB assay showed that BO-modified conjugates decreased the cytotoxicity of PAMAM against both HBMEC and C6 cells and exhibited higher BBB transportation ability than BO-unmodified conjugates; moreover, modification with FA increased the total uptake of DOX by C6 cells and enhanced the cytotoxicity of DOX-polymer against C6 cells. Therefore, FA-BO-PAMAM is a promising nanodrug delivery system in employing PAMAM as a drug carrier and treatment for brain glioma.

Objective:To provide a potential platform for transferring specific antigen against fish bacterial diseases based on attenuated Lm (EGDe-ΔactA/ΔinlB).Methods: Attenuated Lm (EGDe-ΔactA/ΔinlB) was used to express outer membrane protein K (Ompk),a conserved and effective vaccine candidate in vibrio.The identification of recombinant strains and detection of antigen genes were operated with PCR and RT-PCR,respectively.Results: The results of PCR showed that Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.The identity of foreign gene was 100％ compared with sequence of NCBI.The analysis of transcription showed that the expressions of Ompk in L-O,L-L-O and L-P-O were significant (P<0.001).Moreover,the expression of Ompk in the condition of antibiotic was higher than that in the BHI without antibiotic (P<0.05).Conclusion: Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.

Objective To study the time-toxicity and dose-toxicity relationship of hepatotoxicity induced by Paracetamol Tablets (PT),Compound Paracetamol and Amantadine Hydrochloride Tablets (CPAH),Compound Dextromethorphan Hydrobromide Tablets (CDH),and Chaiqin Qingning Capsules (CQC) with single dose in mice.Methods In the Time-Toxicity relationship study,Kunming mice were randomly divided into control,PT,CPAH,CDH,and CQC group,and mice of.each drug administration group were randomly divided into nine subgroups according to the time (1,2,4,8,12,24,48,72 and 96 h after administration) of blood collection.The acetaminophen contents in PT,CPAH,and CDH groups were 425.98 mg/kg,and the dose of CQC group was 3 680.50 mg/kg.In the Dosage-Time relationship study,mice were randomly divided into control,PT,CPAH,CDH,and CQC high,medium and low dose group.The acetaminophen contents of high,medium,and low dose were 266.24,425.98,and 681.57 mg/kg in PT,CPAH,and CDH group,and the dose of CQC group was 1437.70,2300.31,and 3680.50 mg/kg,10 mice in each group,sex in half.Blood was collected 12 h after administration.Animal behavior was observed every day,blood and organs were collected at the corresponding time points,serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),and alkaline phosphatase (ALP) level were detected,and the organs index of spleen and thymus,liver were calculated.Results There were no significant changes of ALT,AST,ALP,and organs index after once ig administration of CQC at dosage of 1437.70 mg/kg to 3680.50 mg/kg in mice.The study on time-toxicity relationship indicated that,after once administration of PT,CPAH,and CDH at 425.98 mg/kg,mice showed toxic symptom such as hypokinesia,dry hair and so on,12 h was the most obvious,24 ～ 72 h disappeared.The level of ALT,AST,and ALP in serum increased and reached to the peak at 12 h and then restored near normality after 72,24,and 24 h in PT,CPAH,and CDH group.Their organ index of liver,spleen and thymus all had no significant changes.The study on the dosage-toxicity relationship indicated that,there were no significant changes of animal behavior,ALT,AST,ALP,and organs index after once ig administration of PT,CPAH,and CDH at 266.24 mg/kg.Obvious liver injury can be induced by the three drugs with dosage of 425.98 to 681.57 mg/kg and the level of ALT,AST,and ALP increased significantly with the increase of dosage.Their liver index increased significantly with dosage of 681.57 mg/kg,but the organs index of spleen,thymus had no significant changes.Conclusion There was no hepatotoxicity after once ig administration of CQC with dosage of 3680.50 mg/kg in mice.Mice were once ig administration ofPT,CPAH,and CDH with a large dose,may induce acute liver injury and show obvious time-toxicity and dose-toxicity relationships.