Objective To produce a hybridoma secreting stable monoclonal antibodies against Aβ22-35 and to develop a detection method for the assay of Aβ. Methods Spleen cells from Balb/cmice immunized with Aβ22-35-KLH were fused with mouse myeloma cells SP2/0. The techniques of immunoprecipitation and western blotting plus ECL were used to investigate the levels of Aβ in the rat brain. Results Two strains of hybridomas (3A8 and 3B2) secreting stable monoclonal antibodies raised against Aβ22-35 were obtained. The subtypes of Aβ22-35 were IgG3. The levels of Aβ in young and older rat brain were 9.8±2.8 and 13.36±2.65 (pmol/12mg brain tissues, x±s), respectively. Conclusion The Aβ22-35 mAb obtained had high titres and specificity. The levels of Aβ in the older rat brain were significantly increased as compared with the young one (P＜0.05).

The method of homogeneity evaluation for active pharmaceutical ingredient (API) spatial distribution in lyophilized product was investigated for the first time with confocal micro-Raman spectroscopy mapping, using pemetrexed disodium for injection as a model drug. Certain areas of the lyophilized product were scanned to obtain Raman spectra. The classical method ("peak clipping" method) was employed for mapping with characteristic Raman peaks of the API and the excipient. Due to the API being finely dispersed in the excipient in lyophilized products, the classical method cannot discriminate between the two ingredients making the distribution homogeneity difficult to evaluate. The "ratio of characteristic peak intensities" method was then utilized. Using this method, the relative intensity of the characteristic Raman peaks of the API to the excipient was applied for mapping and the relative content of API to excipient was calculated for a homogeneity evaluation of the drug distribution. The validation of this method showed a good linear relationship between the relative intensity and the relative content of API to excipient (r² > 0.99), and the precision and recovery were adequate for homogeneity evaluation of API by Raman spectroscopy mapping. Five products of pemetrexed disodium for injection from different manufacturers were tested through Raman maps applying this method and the histograms of relative Raman intensity were also plotted by frequency to help the homogeneity evaluation of drug distribution. The results showed that there were obvious differences in the drug distribution homogeneity from different products, where a more homogeneous API distribution was found in the brand product. This research provides a reliable method for the homogeneity evaluation of API distribution, which facilitates quality evaluation and process optimization of lyophilized products.

Background Application of confocal microscopy in the investigation of ocular surface system in living eye have been greatly extended in two decades.In vivo confocal microscopy allows the evaluation of the normal or pathological tissue at the cellular level.However,seldom study about the morphology of normal in vivo human bulbar conjunetiva under the confocal microscopy was induced. Objective Present study was to analyze the appearance of normal bulbar conjunetiva cells under the in vivo confocal microscopy. Methods Laser scanning confocal microscopy was used to examine the morphological characteristics of the bulbar conjunetiva in 21 eyes of 15 healthy volunteers.The parameters of confocal microscopy were as follows:resolution 1 μm,wave length 670 nm,field range 400 μm×400 μm.The epithelial cell numbers,dendritics cells density and goblet cells density from superior,inferonasal,nasal and temporal bulbar conjunctiva were calculated respectively and compared and imaged by the HRT3 Rostock Cornea Modual. Results The superficial epithelial cells of the bulbar conjunctiva was seen with the small cell nuclei and blur border.The borders of basal epithelial celt were clearly visible without cell nuclei.The presumed goblet cell presented with a large hyperreflective oval-shaped cell with relatively homogeneous brightness,crowed in groups or mainly dispersed.The orfices at the epithelial surface represented the goblet cells,showing some open and expel contents.The dendritic cell appeared to be hyperreflective corpuscular particles with visible processes among conjunetival epithelial cells.A few dense white fiber meshwork was exhibited in conjunctival stroma with the traverse blood vessels containing cellular elements.The superficial and basal epithelial cell densities were 2556±692and 2985±376 cells/mm2 respectively,and overall goblet and mature dendritic cells densities were 77±39 and 26±35 cells/mm2 respectively.Significant differences were proved in globet cell density and dendritic density among different conjunetival zones(P=0.001,P=0. 000),however,the alteration of conjunetival epithelium cells was insignificant in different area(P=0.204,P=0.130).Conclusion Confocal microscopy is a useful tool for the study of morphology of human bulbar conjunctiva cells in vivo.which offer a valuable aid in the diagnosis of ocular surface disease.

Objective To retrospectively evaluate the mammographic features of breast ductal carcinoma in situ (DCIS)and DCIS with small invasive foci,and to analyze the correlation between the mammographic findings and the prognostic biologic factors.Methods The mammographic examination was performed in 95 consecutive women with breast DCIS(n = 50)and DCIS with invasive foci(n = 45 ).The prognostic biologic factors including progesterone receptor(PR),C-erbB-2,and p53 were evaluated in 62 of 95 cases.Categorical data were expressed as percentages and analyzed by using the X~2 test,and furthermore the odds ratio was measured.Results(1)Only one abnormality was seen on mammography in 62 patients. Combined two abnormalities on mammography were seen in 26 patients.Mammograms were normal in 7 patients.(2)Calcifications with or without other abnormality were noted in 62 cases.Of them,73% (n =45)had higher probability of malignancy calcifications and the others were intermediate concern calcifications.Clustered calcifications(36 lesions)was the most common distribution,which usually accompanied by another abnormality.And then were segmental(18 lesions)distributed pattern.As far as the shape of mass (n = 22)was concerned,the oval shaped lesion(13 cases)was the most common,and the margin of the mass appeared as ill-defined in 15 eases,microlobulated in 1,circumscribed in 4,and obscured in 2,respectively.Isodensity mass had a higher frequency in this group(12/22,55%).Other non-calcification findings included architecture distortion(7 cases),local asymmetry (15 cases),global asymmetry (5 cases),and solitary dilated duct (3 cases),and most of them accompanied with other signs. (3)For expression profile of the biological factors,significant differences were found among malignant calcification group,intermediate concern calcification group,and non-calcification group. The odds of PR positive for the lesions noted as non-calcification were 11.00 times higher (X~2 =8.571 ,P=0.003 ;95% CI, 1.998—60.572)than the lesions noted as intermediate concern calcifications,and 8.80 times higher (X~2 = 9.748,P=0.002 ;95% CI,2.024—38.253)than the lesions noted as malignant calcifications.The odds of C-erbB-2 positive for the lesions showed as malignant calcifications were 12.35 times higher (X~2=7.353, P=0.007 ;95% CI,1.447—105.443)than the lesions showed as non-calcification,and 5.74 times higher (X~2=4.977,P = 0.026;95% CI,1.110—29.645)than the lesions showed as intermediate concern calcifications.Conclusion The mammographic features of DCIS and DCIS with small invasive foci were characteristic.Mammographic findings could be a prognostic markers,which could provide a possibility for making a treatment plan.

Objective To evaluate the follow-up results of patients with cervical intraepithelial neoplasia Ⅲ(CIN Ⅲ)after surgical treatment.Methods A retrospective analysis of consecutive patients with CIN Ⅲ after surgical treatment between Jan 1st,1999 and Jun 30 th,2004 was performed.The follow- ups of the patients after surgical treatment were assessed.Results In the follow-up of patients with CINⅢ after surgical treatment,the rate of abnormal cytology was 9.3%.The rate of follow-up was higher in patients with cervical conization than in patients with initial hysterectomy and in patients of the oncological group than of the non-oncological group.The rate of follow-up was lower in patients over 40 years old.There was no difference in the residential areas of the patients.Conclusions Cytological follow-up of patients with CINⅢ after operation is varied.The rate of follow-up is lower in patients over 40 years old,in patients having initial hysterectomy and in patients of the non-oncological group.The rate of follow-up is associated with the knowledge about CIN of both surgeons and patients.

Objective To study the mental health status of nursing students from 2010 grade and its influencing factors so as to explore an effective way to mainrain their psychological health and enhance psychological education.Methods 153 nursing freshmen(female)from 2010 grade of two Universities in Jilin were assessed by using University Personality Inventory(UPI)and self-made questionnaire,and the results were statistically analyzed with SPSS 12.0 software.Results There were three kinds of problems prevailing among the students,7.33%(type 1),25.33%(type 2),and 67.33%(type 3),respectively.Conclusions The psychological problems of nursing freshmen exist to varying degrees,which should call the attention of college student workers and psychological education workers.

OBJECTIVETo observe the intervention and mechanism of Qushi Huayu Recipe (QHR) on gene expression profiles in high lipid diet induced fatty liver rats.

METHODSFatty liver model was prepared in 20 male SD rats using single high fat diet (88% common forage +2% cholesterol +10% lard). Four weeks after modeling they were divided into the model group and the QHR group according to random digit table, 10 in each group. QHR (at 0. 93 g crude drug/100 g body weight) and distilled water was respectively to rats in the QHR group and the model group by gastrogavage while modeling, once per day. Meanwhile, 10 SD male rats were recruited in a normal group, administered with equal volume of distilled water by gastrogavage. At the end of week 8 all rats were sacrificed, and blood and livers were collected for subsequent analysis. Contents of liver triglyceride (TG) and free fatty acid (FFA) , activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using biochemical assay. Pathological changes of liver tissue were observed using H&E and oil red O stain. Liver gene expressions were detected by Affymetrix gene expression profiles. Differentially expressed genes were compared between the QHR group and the model group, functions of differentially expressed genes and signal pathways involved analyzed. Ten differentially expressed genes involved in glycolipid metabolism with fold change more than 2 were selected for verification by real-time PCR.

RESULTS(1) Compared with the normal group, contents of liver TG and FFA, and serum activities of ALT and AST obviously increased in the model group (P <0. 01). Compared with the model group, contents of liver TG and FFA, and activities of ALT and AST obviously decreased in the QHR group (P <0. 05, P <0. 01). QHR could reduce high fat induced fatty degeneration of liver cells , alleviate inflammation, and improve pathological changes of liver tissue. (2) Compared with the model group, there were 80 differentially expressed genes (with fold change > 2, P < 0.05) with clear functions and appointed gene names, including 44 up-regulated and 36 down-regulated genes. Eighty genes were involved in 27 signal pathways with statistical difference, including glycerolipid metabolism, adipocytokine signaling pathway, insulin signal pathway, drug metabolism signal pathway, etc (P < 0.05). (3) RT-PCR results of 10 glycolipids metabolism regulating genes such as Gk, Scd1, Gpat2, G6pc, Irs1, and so on showed that all RT-PCR genes were completely coincide with up-regulated or down-regulated tendency in results of gene chips. 80% genes had approximate fold change.

CONCLUSIONQHR could regulate gene expressions related to fat metabolism, carbohydrate metabolism, anti-lipid peroxidation, and drug metabolism in high fat diet induced fatty liver rats, and its comprehensive pharmacological actions could be manifested.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Carbohydrate Metabolism ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Fatty Acids, Nonesterified ; metabolism ; Fatty Liver ; metabolism ; Lipid Metabolism ; Lipid Peroxidation ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transcriptome ; drug effects ; Triglycerides ; metabolism

AIM:To remark the effect of Qingguang'an Ⅱ on expression of PAX6, Ngn1, and Ngn2 mRNA of rats with chronic high intraocular pressure.METHODS:Totally 40 male SD rats were randomly divided into 6 groups, that was:A:blank group, B:model group, C:Qingguang'an Ⅱ low dose group, D:Qingguang'an Ⅱ moderate dose group, E:Qingguang'an Ⅱ high dose group, F:Yimaikang disket group.B, C, D, E, F groups of experimental rats were established the model of chronic high intraocular pressure (IOP) by cauterizing of superficial scleral vein.Animal model was established successfully by using monitoring IOP consistently keep above 25mmHg for 8wk as cut-off criterion.Tissues of Eyes were obtained after intragastric administration for 2wk and 4wk.The expressions of PAX6, Ngn1, and Ngn2 mRNA were investigated by Real-time PCR.RESULTS:At the time-point of 2wk, PAX6, Ngn1, and Ngn2 mRNA in group B were statistically expressed in lower level comparing with other groups (P<0.05).Moreover, at the time-point of 4wk, PAX6, Ngn1, and Ngn2 mRNA in group C, D and E were statistically expressed in higher level comparing with group F (P<0.05).Besides, PAX6, Ngn1, and Ngn2 mRNA in group C and D were statistically expressed in lower level comparing with group E (P<0.05).PAX6, Ngn1, and Ngn2 mRNA in group C and D were expressed in similar level(P>0.05).CONCLUSION:In summar, Qingguang'an Ⅱ and Yimaikang disket can remarkably increase the expressions of PAX6, Ngn1, and Ngn2, which suggest protecting the optic nerve of rats caused by chronic high IOP.What's more, this study indicated that, in the protection of optic nerve of rats with chronic high IOP, the high dose of Qingguang'an Ⅱ at the time-point of 4wk was the better choice.

Objective:To detect the change of proportions of αβT lymphocytes in mesenteric lymph nodes of miR-7 knockdown (miR-7KD)mice,and preliminarily explore its importance.Methods: The volume,weight index and total cells number of mesenteric lymph node in miR-7KD mice were measured.The pathologic morphology change of mesenteric lymph nodes was observed by HE stai -ning.And the changes on proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice were analyzed by Flow cytometry.Results:Compared with those of WT (wild type) mice,the volume,weight index,and the total cells number of mesenteric lymph node were significantly increased ( P<0.05 ).Moreover ,the pathologic morphology was significantly changed.The proportion and numbers of T lymphocyte were significantly increased;however,the proportion of B lymphocyte were significantly decreased (P<0.05). Notably,the proportion and number of CD4+T cells and CD8+T cells were significantly increased (P<0.05).Meanwhile,CD62L+T cell proportion were vigorously reduced and CD 69+T cell and IFN-γ+T cell proportion were notably up-regulated,which belong to CD4+and CD8+T cell(P<0.05).Conclusion:There was significant influence in the proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice,suggesting that miR-7 might play an important role in the composition and function of lymphocytes in mesenteric lymph nodes.

Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.