10.3969/j.issn.2095-4344.2013.24.024

Objective To detect the expression of BP1 gene in thyroid cancer and its relationship with clinicopathological features of thyroid cancer.Methods BP1 gene expression in 60 cases of thyroid cancer tissues and 20 cases of normal thyroid tissues were detected by in situ hybridization and immunohistochemistry.Results The positive expression rate of BP1 mRNA was 78.3 ％ (47/60) in the 60 cases of thyroid cancer tissues while it was 20％ (4/20) in the 20 cases of normal thyroid tissues detected by in situ hybridization.The difference had statistical significance (P ＜ 0.05).Of the 3 pathological types of thyroid cancer,the positive expression rate of papillary carcinoma was 81.6％ (40/49),follicular carcinoma 85.7％ (6/7),and medullary carcinoma 25.0％ (1/4).The expression of BP1 mRNA had statistical difference between medullary carcinoma and other pathological types like papillary carcinoma and follicular carcinoma (P ＜ 0.05).The positive expression rate of BP1 protein was 93.3％ (56/60)in the 60 cases of thyroid cancer tissues while it was 10.0％ (2/20) in the 20 cases of normal thyroid tissues detected by immunohistochemistry.The difference had statistical significance(P ＜0.05).Conclusion BP1 gene expression is up-regulated in human thyroid cancer and it is related to tumor stage and pathological type but not related to patients' age,sex or lymph node metastasis.

A strain of canine parvovirus (CPV) was isolated from feces of an ill puppy in an animal hospital in Wuhan, China. It was designated as CPV/WH02/06. This isolate was identified as serotype CPV-2a by the hemagglutination test, CPV Ag detection strip, electron microscopy, and PCR. The vp2 gene was cloned and sequenced and assigned GenBank accession number EU377537. A 1242 bp segment of the 5' region of the vp2 gene was cloned and inserted into the binary vector pBI121 and used for Agrobacterium-mediated tobacco transformation. Transgenic tobacco plants were selected on MS medium supplemented with 100 μg/mL kanamycin and 100 μg/mL timentin. Integration of the vp2 gene into the tobacco genome was confirmed by PCR using T1 progeny plants, and the expression of the VP2 protein was confirmed by Western blotting.

AIM: To investigate the role of epigenetic modification in Pdx-1 gene transcription and expression, and to compare the differences between epigenetic modifications of Pdx-1 gene promoter in various cell types of mice. METHODS: The promoter DNA methylation and histone modification status of Pdx-1 and MLH1 genes in NIT-1 cells, NIH3T3 cells and mouse embryonic stem cells were measured by chromatin immunoprecipitation-real time PCR method. The expression levels of these genes in the three cell lines were measured by real time RT-PCR. The relation between epigenetic modifications and gene expression was analyzed. RESULTS: (1) Compared to mES cells, there was lower DNA methylation and higher H3K4m3 modification levels in the promoter of Pdx-1 gene in NIT-1 cells (P<0.05). DNA methylation, H3 acetylation, H3K4m3 and H3K9m3 modification levels in the promoter of Pdx-1 gene in NIH3T3 cells were distinctly increased (P<0.05). (2) Pdx-1 gene transcription expressed only in NIT-1 cells. The Spearman's rho between Pdx-1 gene expression and DNA methylation (r=-0.802,P<0.01) was observed. The Pearson correlation between Pdx-1 gene expression and H3K4m3 modification (r=0.997,P<0.01) was also found. The Spearman's rho between Pdx-1 gene expression and H3K9m3 modification (r=-0.879,P<0.01) was observed. (3) No correlation between housekeeper MLH1 gene expression and epigenetic modification was found. CONCLUSION: DNA methylation, H3K4m3 and H3K9m3 modification coordinated participate to regulate and control the expression of Pdx-1 gene. It is of great significance to the differentiation of β cells from ES cells.

Objective To investigate the therapeutic effects and mechanism of biliopancreatic diversion (BPD) surgery on type 2 diabetes mellitus(T2DM) in GK rats.Methods 16 GK rats were randomly divided into 2 groups:BPD surgery group included 10 rats undergoing BPD surgery,sham-BPD group included 6 rats undergoing a sham operation.Fasting plasma glucose,insulin,glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide(GIP)were detected one week before BPD surgery and the 1st week,4th week,10th week,26th week after BPD surgery.Oral glucose tolerance test(OGTT) and insulin tolerance test(ITT) were done in the 10th week after BPD surgery.Results There was no statistical difference in fasting plasma glucose,insulin,plasma GLP-1 or GIP between the 2 groups before surgery.Plasma glucose had significant reduction in BPD group compared to that in the sham group(P ＜0.05) and insulin level had no significant difference between the 2 groups.Rats in BPD group had significant improvement in glucose tolerance and insulin sensitivity compared to those in the sham group.Serum level of GLP-1 was significantly elevated in BPD group compared to that before surgery (P =0.0337 at the 1st week after surgery; P =0.0002 at the 4th week after surgery,P ＜ 0.0001 at the 10th week after surgery,P ＜0.0001 at the 26th week after surgery) and that in sham-BPD group(P =0.0354 at the 1st week after surgery,P =0.0032 at the 4th week after surgery,P =0.0001 at the 10th week after surgery,P ＜0.0001 at the 26th week).Serum level of GIP was significantly lowered in BPD group compared to that before surgery(P =0.0189 at the 1st week after surgery; P =0.0007 at the 4th week after surgery,P =0.0003 at the 10th week after surgery,P ＜0.0001 at the 26th week after surgery) and that in sham-BPD group(P =0.0089 at the 1st week after surgery,P =0.0002 at the 4th week after surgery,P =0.0006 at the 10th week after surgery,P ＜0.0001 at the 26th week after surgery).The difference had statistical significance (P ＜0.05).Conclusion BPD surgery can significantly reduce fasting plasma glucose,improve glucose tolerance and insulin sensitivity.The change of serum levels of GLP-1 and GIP may play the major role in BPD treatment of diabetes mellitus.

Aim To study the change of blood pressure in 1079 workers for fifteen years and the effect of anti-hypertension drugs therapy. Methods We investigated the blood pressure of 1079 workers who were enrolled in six organizations in the year 1983 and 1998 respectively. Results During the 15 years: (1)Mean value of blood pressure: systolic blood pressure increased 22 mmHg in man and 16.9 mmHg in women; diastolic blood pressure increased 9 mmHg in man and 12.7 mmHg in women (P<0.05);(2)The incidence of hypertension increased by 25.03% in man and 28.28% in women;(3)The prevalence rate of hypertension is 27.9 percent in people with initially normal blood pressure (1.86%/y) and 72.6 percent in initially broder line hypertension (4.84%/y);(4)The control rate of hypertension is 2.9 percent;(5)The incidence of stroke is highly related to hypertension (P<0.001);(6)46.6 percent patients had a regular drug therapy, mainly reserpini complex(25.2%);(7)Drug therapy has no obviously effect of the control of hypertension and the incidence of stroke. Conclusion Both the mean value of BP and the incidence of hypertension were increased with age. The control rate of hypertension was low and the drug therapy shows little advantage. We should do mach more works to popularize the knowledge of the prevention of hypertension. Improve people's self-prevention. Regular cheek should be given to the hypertension patients.

Objective To investigate the risk factors of electrocoagulation syndrome after endoscopic submucosal dissection (ESD) in patients with colorectal lesions.Methods Clinical data of 145 patients with colorectal mucosal lesions undergoing ESD in People's Hospital of Wuhan University between September 2010 and September 2015 were retrospectively studied.Results Among 45 patients,post endoscopic submucosal dissection electrocoagulation syndrome (PEECS) was developed in 32 cases (22％).The median age in PEECS group was higher (t =-5.783,P =0.000),the median lesion size was larger(t =-5.590,P =0.000),the median length of hospital stay was longer (t =-6.841,P =0.000) than those in non-PEECS group.Univariate regression analysis showed PEECS was associated with the age,lesion size,lesion location,length of hospital stay,malignant tumor,polyps type,resection modality.Multivariable logistic regression analysis showed that the independent risk factors for the development of electrocoagulation syndrome were age ＞65 year (OR =1.123,95％ CI:1.013-1.244,P =0.027),lesion size ＞ 3.5 cm (OR =1.173,95％ CI:1.015-1.357,P =0.031),malignant tumor (OR =3.498,95 ％ CI:1.460-8.379,P =0.005),hospital stay ＞ 10 d (OR =2.480,95％ CI:1.346-4.569,P =0.004),non-rectal lesions (OR =12.612,95％ CI:3.446-46.157,P =0.000).Conclusion Attention should be paid for colorectal lesion patients with high risk of PEECS,when endoscopic submucosal dissection is performed.

Background The study on the classification of fungi is very important for the diagnosis and treatment of fungal keratitis.Identifying the different species of filamentous fungi is a critical factor for the application of anti-fungal drug in treating keratitis.ObjectiveThis report studies the relationship between the genotype of filamentous fungi and the clinical factors.MethodsFifty-two patients with filamentous fungal keratitis determined by clinical and laboratory examination were recruited in Tongren Hospital from January 2006-December 2006.The lesions were graded on the severity of the corneal ulcer and the presence of hypopyon.The filamentous fungal keratitis was treated with topical and systemic administration of anti-fungal drugs or corneal transplantation.The isolates were cultured in potato culture and identified by morphological characteristics based on the Nelson criterion and genotyped by the rDNA ITS method.The clinical data was retrospectively analyzed.ResultsForty-eight species (eubacteria are bacteria,not fungi)of fungus were identified by morphological characteristics,and the filamentous fungi were divided into 4 types based on the phylogenetic relationships within the rDNA ITS of the 52 filamentous fungi.The morphological characteristics and genotype were confirmed in 48 strains of eubacteria and 31 strains of 52 filamentous fungi (90.3%).The 4 groups of fungi were classified by genotype as follows:group 1 represents 22 strains including 20 strains of Fusarium solani and 2 strains of Fusarium oxysporum;group 2 represents 12 strains including 8 strains of Fusarium moniliformis,3 strains of Fusarium proliferatum and 1 strain of Fusarium incarnatum;group 3 represents 5 strains including 1 strain of Fusarium moniliformis and 4 unknown strains;group 4 represents 13 strains including 10 strains of Aspergillus spp.and 3 strains of Alternaria spp.Significant differences were found in the disease duration (P=0.00),inducing cause (P=0.03),ulcer grade (P=0.01)and outcome of the anti-fungal treatment (P=0.035)when compared between group 1 and 2 with group 3 and 4.Conclusion Filamentous fungi that cause keratitis could be correctly identified by sequencing the internal tanscribed spacer of rDNA.There are significant clinical differences among the groups classified by genotype.

Objective To investigate the clinical manifestation of trachoma,and to select the appropriate laboratory test for clini- cal diagnosis.Design Retrospective case series.Participants Retrospective analysis of medical records from 61 patients with trachoma from Jan 2003 to Aug 2006 in Bejing Tongren Eye Center.Methods Grades of trachoma diagnosis were according to the criteria de- signed by Chinese Ophthalmological Society (1979).The general state of health,case history,and the laboratory investigations of pa- tients were recorded.Laboratory tests included the conjunctiva scraping for inclusions,C.trachomatis immune antigen test and PCR test.Main Outcome Measures Manifestation of corneal and conjunctiva,the results of laboratory tests of C.trachomatis.Results Out of sixty-one patients including 28 males and 33 females,the average old was (29.05?19.99) years.88.5% cases were inⅠstage of tra- choma,8.2% were inⅡstage,and 3.3% were inⅢstage.The C.trachomatis inclusions were found in 7 (11.5%) scraping smears.42 (68.9%) cases were positive in C.trachomatis antigen test.46 (75.4%) cases were positive in PCR tests.The positive rates of antigen and PCR test were significantly higher than that of scraping (P=0.00).Conclusions A majority of clinical patients were inⅠstage of trachoma.The degree of their distress was minimal.It is necessary to apply C.trachomatis,antigen test or PCR test to improve the clin- ical diagnosis.

Objective To provide the evidences for the management strategies of ventilator-associated pneumonia (VAP) in neonates,we systematically reviewed all related studies and analyzed the high-risk primary disease and medical factors of VAP in neonates.Methods We retrieved all related studies in CNKI,Wanfang,VIP,CBM,Pubmed and Embase and evaluated their quality by Newcastle-Ottawa Scale and analyzed all data by qualitative and Meta-analysis.Results There were 12 case-control studies with higher methodological quality and involving 1 994 neonates and with 708 VAP patients.Six studies involving 872 neonates were included,the odds ratio of respiratory distress syndrome(OR=2.81) and malnutrition(OR=5.18) had significant differences between VAP and non-VAP group.Seven studies involving 1 110 neonates were included and the odds ratio of patients with corticosteroids (OR=3.12),central inhibitors (OR=2.31),antacids (OR=4.35) and Gamma globulin with large doses (OR=2.35) had significant differences between VAP group and non VAP.Four studies involving 554 neonates were included and the odds ratio of patients with closed chest drainage (OR=1.81)and umbilical vein catheterization (OR=9.19) had significant differences between VAP group and non VAP.Six studies involving 1 139 neonates were included and the odds ratio of patients with parenteral nutrition (OR=1.82)and blood transfusions (OR=2.49) had significant differences between VAP group and non VAP.Conclusions Our study confirms that the respiratory distress syndrome and malnutrition corticosteroids,central inhibitors,antacids,Gamma globulin with large doses,closed chest drainage,umbilical vein catheterization,parenteral nutrition and blood transfusions are important risk and early-warning factors.