AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet.
METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods.
RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the
model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon.
CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.

Tumor genesis and development often result from deregulation of important biological pathways at the gene expression level. Although there has been much work focused on searching gene sets using gene expression data or other prior information, proper statistical testing of the gene sets is still an open question. Most studies have expanded the testing method of a single gene into the gene sets. Parametric statistical analysis of gene sets ( p-SAGE ) was presented for determining the significant gene sets or pathways associated with a phenotype of interest. The method was applied to brain tumor experiments to identify many gene sets. Some of the newly discovered gene sets were related to signal transduction and immunity. This simple and effective method gives useful biologically meaningful results.

To study the protective effect of Arctigenin in goto-kakizaki (GK) rats combined with hypertension macroangiopathy. Six-week-old GK rats were divided randomly according to blood glucose level into four groups: the model group and low, middle and high dose arctigenin groups (12.5, 25, 50 mg x kg(-1)), with Wistar rats as the normal group. All of GK rats were given high-glucose and high-fat diet. After 16 weeks, GK rats were orally administrated with 10 mg x kg(-1) x d(-1) N-Ω-nitro-L-arginine methyl ester for eight weeks. During the modeling, all of arctigenin groups were orally administrated with different dose of arctigenin twice a day; The model group and the normal group were given solvents. At the beginning, mid-term and end of the experiment, blood glucose was measured. At the end of the experiment, efforts were made to detect blood pressure, collect abdominal aortic blood after anesthesia, fix thoracic aorta after bloodletting to make paraffin sections, observe morphological characteristics and detect the expression of VEGF by immunohistochemistry. According to the results, the blood glucose rose in all GK rats, with no significant difference between the drug group and the model group. At the end of the experiment, the blood pressure significantly increased in GK rats, indicating that Arctigenin could notably reduce the blood pressure in GK rats in a dose-dependent manner. The blood routine test showed increases in both the total white blood cell count and differential blood count, MPV and PDW, abnormal blood platelet parameters and decrease in PLT in GK rats, suggesting that Arctigenin could remarkably reduce the total white blood cell count and differential blood count, MPV and PDW. The thoracic aortic morphological observation revealed obvious endangium lesions in GK rats, demonstrating that Arctigenin could ameliorate the lesion extent. VEGF immumohistochemical staining showed a higher VEGF expression in the model group but lower expression in Arctigenin groups. In conclusion, Arctigenin had a protective effect on aorta in GK rats. Its mechanism may be related to blood pressure lowering, anti-inflammation, improvement in blood platelet function and reduction of VEGF expression.
Animals ; Blood Glucose ; metabolism ; Blood Pressure ; drug effects ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; physiopathology ; Diabetic Angiopathies ; etiology ; metabolism ; physiopathology ; prevention & control ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Furans ; administration & dosage ; Humans ; Hypertension ; etiology ; metabolism ; physiopathology ; prevention & control ; Lignans ; administration & dosage ; Male ; Rats ; Rats, Wistar

OBJECTIVE:To establish the quality standard of Liyan mixture. METHODS:TLC was conducted to identify the Scrophularia ningpoensis,Terminalia chebula and Glycyrrhiza uralensis;HPLC was conducted to determine the content of chloro-genic acid. The column was Agilent Eclipse Plus C18 with mobile phase of methanol-0.2% phosphoric acid (18:82,V/V) at flow rate of 1.0 ml/min,the detection wavelength was 327 nm,column temperature was 30 ℃ and the injection volume was 20 μl. RE-SULTS:TLC showed S. ningpoensis,T. chebula and G. uralensis had clear spots and good separation. There was no interference for negative control. The linear range of chlorogenic acid was 0.16-1.6 μg(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 1.0%,recovery was 96.9%-101.4%(RSD=1.17%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Liyan mixture.

Background and purpose:The expression of ERβin triple negative breast cancer(TNBC) might be associated with good prognosis in TNBC patients. ERβand ERαhave considerable homology. FOXA1 plays an important role in ERαexpression and function. The aim of this study was to analyze the expression of FOXA1 and ERβin TNBC and the relationship between them and the clinicopathologic characteristics and prognosis. Methods:The breast cancer samples in Peking Union Medical College Hospital were collected from Nov. in 2011 to Dec. in 2013, and TNBC were screened out based on the expression of ERα, PR and HER-2. Thirty ERβ-negative samples and 30 ERβ-positive samples were selected randomly according to the ERβexpression. We used immunohistochemical method to detect the expression of FOXA1. Finally, 48 TNBC samples were obtained to analyze the results. Results:The total positive rate of FOXA1 was 35.4%(17/48). In the ERβ-positive group, the positive rate of FOXA1 was 35.7%(10/28),and in the ERβ-negative group, the positive rate of FOXA1 was 35% (7/20). The expression of FOXA1 in these 2 groups had no signiifcant difference (P=0.83), which indicated that there was no relation between ERβand FOXA1. The FOXA1 positive group and FOXA1 negative group also showed no signiifcant difference in age, tumor size, and lymphatic metastasis number in axilla, tumor grade, tumor stage, NPI and DFS. However, Ki-67 showed negative correlation with FOXA1 expression (P<0.01). Conclusion:FOXA1 expression had no relationship with ERβexpression in TNBC. Ki-67 showed negative correlation with FOXA1 expression, which might hint that the proliferation of tumor cell was lower in FOXA1 positive TNBC.

Objective To summarize the clinical characteristics and causes of ischemic cerebrovascular disease(ICD)in children.Methods A retrospective analysis was conducted on the clinical data of 53 cases with ICD from Feb.2002 to Jun.2008 at the department of neurology in Wuhan Children's Hospital.The self-designed questionnaire of children with ICD was used,whose items included patients' age,gender,personal history,clinical features,cerebrospinal fluid examination,neurological imaging,immunologic examination,metabolic examination,and so on.Results Of 53 children with ICD,30 cases(56.6%)were male,and 23 cases(43.4%)were female.Patients' age varied from 9 months to 12 years old,in which 45 cases(84.9%)were less than 6 years old.Patients from rural area(60.4%)were more than those from city(39.6%).Ratio of limb paralysis was 75.5%(40 cases)in first clinical symptomatology of children with ICD,including hemiplegia in 32 cases(60.4%),alternate hemiplegia in 5 cases(9.4%)and monoplegia in 3 cases(5.7%).Skull CT/MRI scan was performed to reveal 27 cases(50.9%)with basal ganglia region infarction and secondly 15 cases(28.3%)with multi-lobar infarction.Forty cases were found in abnormal cerebrovascular image by means of magnetic resonance angiography/digital subtraction angiography,in which middle cerebral artery and its branches were involved in 21 cases(52.5%).There were 41 cases(77.4%)of patients to be found with clear causes,of which 13 cases(24.5%)were of infections,8 cases(15.1%)of moyamoya disease,5 cases(9.4%)of cerebral vascular malformations,4 cases(7.5%)of head trauma.However,another 12 cases(22.6%)of patients had unknown etiology.Conclusions Children with ICD had characteristics themselves.The limb paralysis was mostly the first symptoms,and the middle cerebral artery and its branches lesions were the most common locations in children with ICD,and next the internal carotid artery involvement,anterior cerebral artery involvement,posterior cerebral artery involvement,cerebral vascular malformations,and so on.Their major cause was infection,followed by Moyamoya disease,cerebrovascular malformations and head trauma,and there were still some unknown causes.

As the main part of the teaching activities,students play an important role in the teaching reform.The students were trained through 3 pathways,“Extending teaching activities from the classroom to the outside”,“Development from basic to clinical knoledge” and “Culturing students' innovative consciousness”,so as to allow them to give full play in teaching reform,to enhance their ability of practice and learning by themselves,to culture their innovative consciousness and to develop students' leading functions in the anatomy teaching reform.

Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.

Objective To study effect and mechanism of fungus polysaccharide PSM-a of Polyporus sp.M05 on S180 bearing mice. Methods MTT method was used to detect the inhibiting role of PSM-a on S180 cells proliferation in vitro. S180 mice model was established,and was administered by gavage. Tumor volume was detected, and the ratio of tumor to mile weight and inhibiting tumor rate. The activity of NK and LAK cells on the target cells was analyzed by MTT colorimetric assay ;HE stain was used to detect the necrosis of tumor cells. Results PSM-a could inhibit S180 cells grouth in vitro. PSM-a could decrease the tumor weight and increase the ratio of tumor volume and mice weight; Tumor inhibiting rate reached 80% and above when treated with 250 μg/nml PSM-a. PSM-a could increase the activity of NK and LAK cells, and necrosis happened. Conclusion PSM-a could significantly inhibit the growth of S180 cells, and the mechanism bnay be related with the increased killing activity of immunne cells to tumor cells.

Objective To investigate the effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA. Methods After establishing the in vitro differentiation model with HMBA on K562 cells, the MTT assay was used to detect the proliferation of K562 cells, the cell cycle profile was detected by flow cytometry, and the mRNA expression of TGF-β1, SMAD3, SMAD4 and EVI1 was measured by RT-PCR assay. Results HMBA could inhibit the proliferation and promote the differentiation of K562 cells obviously, which was time and concentration-dependent, and the 72 h corresponding IC50, was about 2 mmol/L. Within 72 h, flow cytometry assay indicated that the ration of G0-G1 phase cells was up-regulated, and the results of RT-PCR showed that relative mRNA expression of TGF-β1, SMAD3 and SMAD4 at mRNA level was increased gradually while that of EVI1 was decreased gradually. Conclusion HMBA can inhibit K562 cells proliferation through TGF-β1/SMAD signaling pathway.