Objective To investigate the expression of nuclear factor-κB in alveolar macrophages of paraquat-induced rats and the effect of diallyl sulfide on it.Methods Forty five male wistar rats were randomly divided into three groups,namely control group,model group,and DAS treatment group (n =15 in each).The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by intra-gastric administration,while the equal volume of normal saline (NS) was given to the rats in control group instead.The dose of 100 mg/kg of DAS was given to rats by intra-peritoneal injection in DAS treatment group.The equal volume of NS was given to the rats by intra-peritoneal injection in model group and control group instead.The rats of model group and DSA treatment group were exposed to paraquat once a day for 14 days.Five rats in each group were sacrificed at 3,7,14 days.Alveolar maerophages were harvested by bronchalveolar lavage (BAL).The protein content of BAL fluid were examined.The exprossion of NF-κB was measured with immunocytochemistry technique.Results Alveolar macrophage cultures were carried out by using differential adherence of isolated and purified alveolar macrophages,and after 30 minutes culture,more adherent macrophages can be seen.Compared with model group,the protein content of BAL fluid at dfferent intervals in the control group were obviously lower,especially on the 3 rd day (261.6 ± 17.16) μg/mL vs.(673.4 ± 151.9) μg/mL;7 d (265.6 ± 18.37) μg/mL vs.(581.3 ± 134.58) μg/mL;14 d (253.8 ± 11.43) μg/mL vs.(589.07 ± 33.85) μg/mL,P ＜ 0.05.Comparisons of protein content in BLA fluid between PQ group and DAS treatment group were on the 3 rd day (673.4 ± 151.9) μg/mL vs.(342.9 ±39.03) μg/mL;on the 7 th day (581.3 ± 134.58) μg/mL vs.(383.7 ±7.37) μg/mL,P＜0.05;on the 14 th day (589.07±33.85) μg/mL vs.(282.9±15.59) μg/mL,P＜0.05.The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell cytoplasm in the control group,while high NF-κBp65 expression was found in nuclear in the model group.Minimal NF-κBp65 expression was found in the cell cytoplasm in the DAS treatment group,and integral A value was significantly lower in the DAS treatment groups than that in the model group (P ＜ 0.05).Conclusions Treatment with an intra-peritoneal injection of DAS is capable of attenuating the extent of PQ-induced ALI in rats by lowering BLA fluid protein content,inhibiting the expression of NF-κB in alveolar maerophages.

Taking children malnutrition,diabetes insipidus and diarrhea as examples,it introduces doctor He Guantao’s clinical experience in treating children disease with ebony,viewing that he makes conclusion with practice,often curing hard diseases with smart formula descriptions.

Objective To explore the Cep70 by adjusting the stability of acetylated alpha tubulin,participate in breast cancer drug resistance mechanisms.Methods (1) In order to induce taxol drug resistance cell line Michigan cancer foundation-7 (MCF-7)/pac,high-dose shock treatments taxol MCF-7 was used for 6 months,until the cells can grow in 3.5 μmol/L of paclitaxel.(2) The 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method was used to detect inhibition rate by taxol to MCF-7 and MCF-7/pac cell.(3) Immunofluorescence and Western blot were used to test acetylated alpha-tubulin and Cep70 expression levels in MCF-7 and MCF-7/pac cells.(4) Chemical intervention was used to acetylate apha-tubulin expression,Western blot and polymerase chain reaction (PCR) were used to detect the change of acetylated alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac groups.Flow cytometry and Western blot were used to detect the change of cell cycle.Results (1) IC50 of MCF-7 and MCF-7/pac was 22.47 μ mol/L and 31.38 μmol/L,respectively.(2) Immunofluorescence and Western blot results showed that the expression of acetylation of alpha-tubulin in resistant MCF-7 cell/pac was obviously decreased.(3) Real time polymerase chain reaction (RT-PCR) and Western blot showed Cep70 expression was consistent of acetylation of alpha-tubulin.(4) After incubation with paclitaxel for 24 hours,the expressions of acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac were increased,but the extent of MCF-7 cell was much higher.Instead,incubation with nocodazole after 24 hours,the acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac cells were obviously lowered.(5) After paclitaxel intervention,compared to the same group MCF-7 cells,the G2 phase ratio in MCF-7/pac cells was lower.In addition,given nocodazole after the intervention,compared to the same group MCF-7 cells,the ratio of G2 phase in MCF-7 cell/pac was significantly decreased.Conclusions Cep70 decreased the expression of the acetylated alpha-tubulin,reduced the stability of microtubules,which could be an important mechanism of taxol drug resistance.

Objective To reveal acetylated a-tubulin acts as pressure sensors,sensing the changes in extracellular matrix to impact on the invasion and metastasis of breast cancer.Methods The acetylated alpha microtubule protein expressions were detected,and its relationship with invasion and metastasis in breast cancer clinical specimens and different molecular subtypes of breast cancer cell lines were analyzed.The expression of acetylated a-tubulin was interfered through chemical methods,and cell morphology and the change of the invasion and metastasis ability were detected under the condition of different matrix hardness.Results The expression of acetylated a-tubulin was highest in basal-like breast cancer tissue and cell lines,and was lowest expressed in the luminal B breast cancer tissue and the breast cancer cell lines.The expression of acetylated a-microtubule was positively correlated with the occurrence of breast cancer.Under the condition of soft substrate cultivation,cell polarization was declined,becoming the circular or oval shape.The acetylated a-tubulin caused reduction in cell polarity,accompanied with less invasion and metastasis ability.Conclusions The acetylated a-tubulin acts as pressure sensors,sensing the sclerosis of extracellular matrix in the process of tumorigenesis,promoting invasion and metastasis of breast cancer.

OBJECTIVE:To provide reference for rational use of drugs for gastrointestinal ulcer. METHODS:The drug use for gastrointestinal ulcer in outpatient department of our hospital during 2013-2015 was analyzed retrospectively in respects of drug name,consumption sum,DDDs,DDC,single/multiple diseases prescription,therapeutic efficacy,rationality of presoription,etc. RESULTS:A total of 8 kinds of drugs use for gastrointestinal ulcer in outpatient department of our hospital during 2013-2015, among which the consumption sum and DDDs of Omeprazole capsule and Rabeprazole capsule increased rapidly,and their DDC were the higher others;those of other drugs kept stable. The proportion of acid-inhibitory drugs in single disease prescriptions was lower than in multiple diseases prescriptions,while that of antiacid was higher than in multiple diseases prescriptions,with statisti-cal significance (P<0.05). The total effective rate of patients with gastrointestinal ulcer gradually increased from 77.98% to 82.68% in outpatient department of our hospital during 2013-2015,there was statistical significance between 2015 and 2013(P<0.05);the proportion of irrational prescriptions decreased from 5.01% to 2.12% year by year,with statistical significance among those years (P<0.05). CONCLUSIONS:Omeprazole is the first choice for gastrointestinal ulcer in outpatient department of our hospital with good therapeutic efficacy. There still are some irrational drug use in prescriptions,and clinicians should pay attention to those situations.

Objective To develop a TaqMan MGB probe-based,sensitive and specific real-time fluorescence quantitative PCR assay for rapid detection of Helicobacter hepaticus.Methods Primers and probes specific toflaB gene of Helicobacter hepaticus were designed.A TaqMan MGB probe-based,real-time fluorescence quantitative PCR was established.The specificity,sensitivity and stability of the assay were assassed.Then,the established TaqMan MGB probe real-time fluorescence quantitative PCR assay was applied to detect Helicobacter hepaticus in 1081 clinical specimens during 2008-2011,compared with bacterial isolation and culture method and conventional PCR assay.Results The specificity of this established TaqMan MGB probe-based real-time fluorescence quantitative PCR was high and there were no cross-reactivity with Helicobacter pylori,Campylobacter jejuni,Clostridium piliforme,Pasteurella pneumotropica,Escherichia coli,Pseudomonas aeruginosa.The detection limits was 8.3 copies.The correlation coefficient and slope value of standard curve were 0.999 and -3.227,respectively and the efficiency of TaqMan MGB-based probe realtime fluorescence quantitative PCR assay was 100％.The TaqMan MGB-based probe real-time fluorescence quantitative PCR and conventional PCR were preformed to detect Helicobacter hepaticus in 1081 clinical specimens,a total of 86 specimens were positive for Helicobacter hepaticus.However,there was only 4 specimens were positive by bacteria isolation and culture method.The results showed that TaqMan MGB -based probe real-time fluorescence quantitative PCR for Helicobacter hepaticas was more sensitive than bacteria isolation and culture method,and it could detect Helicobacter hepaticus DNA from clinical specimens directly,and detection time is only 2 hours.Conclusion The TaqMan MGB-based probe real-time fluorescence quantitative PCR assay was a reliable,specific,sensitive and useful tool for rapid detection of Helicobacter hepaticus.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

Objective To improve the accuracy of detection through analyzing the phenotypes of P.pneumotropica isolates in laboratory animals in Beijing area .Methods 306 suspicious P.pneumotropica strains were identified by biochemical identification and 16S rDNA sequencing.Then, to obtain the phylogenetic relationships combined with colony characteristics on blood agar plates and biochemical characteristics of 53 biotypes .Results BD Phoenix 100 automated bacterial identification system and 16S rDNA sequencing identified P.pneumotropica positive rate of 306 isolates were 164/306 and 227/306, respectively.There were 140 phenotypes in 227 true-positive strains, of which 106 were biotype Heyl and 23 were biotype Jawetz .Conclusions In the samples of laboratory animals in Beijing area , P.pneumotropica infection mainly are of biotype Heyl , and less is of biotype Jawetz .The phenotypes are diverse and widely distributed .

Fifty children with refractory idiopathic thrombocytopenic purpura (RITP) were treated with rituximab (n =26) or vincristine (n =24).The response rate,adverse reaction and recurrence rate in two groups were compared.The CD19 +/CD20 + B cells in peripheral blood were detected by flow cytometry in 26 patients before and after RITP treatment.The response rates of rituximab group was significantly higher than that of vincristine group ( 69.2％ vs. 37.5％,x2 =9.74,P ＜ 0.01 ). There was no significant difference in the rate of adverse reaction between two groups ( 11.5％ vs.8.3％,x2 =0.62,P ＞ 0.05 ).The recurrence rate of rituximab group ( 22.2％ ) was significantly lower than that of vincristine group (55.6％,x2 =7.24,P ＜ 0.05 ).In rituximab group the platelet count after treatment was significantly higher than that before treatment ( t =12.48,P ＜0.01 ),and the ratio of CD19 +/CD20 + B cells decreased significantly after treatment compare to that before treatment ( t =6.71,P ＜0.05 ).Rituximab is effective in treatment of refractory idiopathic thrombocytopenic purpura in children,which may be associated with decreased B cells in peripheral bloos.

Objective To investigate the distribution pattern of the ADAS-Cog scores among the elderly in Beijing and to evaluate the application of ADAS-Cog in distinguishing patients with mild Alzheimer's disease (AD) and healthy elderly. Methods In total, 1616 healthy elderly (NC), 125 elderly patients with non-AD disorders (ND), and 310 patients with probable AD including 201 patients with mild AD and 109 patients with moderate AD (by NINCDS-ADRDA criteria) were recruited in the study and their cognitive performance was measured by the ADAS-Cog.Results In NC group, those older than 80 years and those with less than 5 years schooling scored highest. There was statistically significant correlation between the total scores of the ADAS-Cog and age and duration of schooling in NC group (F=14.34, 113.27,both P<0.01). No correlation was suggested in the mild AD group. The total score of ADAS-Cog was significantly associated with duration of schooling in both moderate AD and ND groups (F=4.18, 8.72, both P<0.05). The total score of ADAS-Cog differentiated the patients with mild AD from NC healthy elderly with AUC ranging from 0.69 to 0.82 in each subgroup by age, and from 0.75 to 0.88 in subgroups by durations of schooling with the highest AUC of 0.88 in the subgroup having more than 15 years of schooling education. Conclusions The ADAS-Cog score in the healthy elderly is significantly associated with age and duration of schooling education. The ADAS-Cog can be applied in the cognitive assessment of Chinese AD patients. The total score of ADAS-Cog could efficiently differentiate patients with mild AD from healthy elderly, especially in subjects with higher education level.