Immunizing mice using different combination antigens of the PI-2a fimbria subunit of Streptococcus agalactiae.

BACKGROUND: Streptococcus agalactiae is the main causal pathogen of bovine mastitis (BM), causing considerable economic loss to the dairy industry worldwide. Vaccines against S . agalactiae play an important role in preventing disease. AIMS: The aim of this study was to evaluate the immunoprotection of S. agalactiae pilus island fusion proteins, ancillary protein 1-ancillary protein 2 (AP1-AP2), ancillary protein 1-bone protein (AP1-BP), bone protein-ancillary protein 2 (BP-AP2), and ancillary protein 1-bone protein-ancillary protein 2 (AP1-BP-AP2) in Balb/c mice. METHODS: Four kinds of fusion antigens and the same volume of Freund's complete adjuvant were mixed vigorously to prepare fusion antigen immuno-samples. The mice were immunized 4 times (on the 0th, 7th, 14th, and 28th days) with these samples with an immunizing dose of 50 g/mouse. After the 4th immunization, serology tests were used to evaluate the antibody. The antibody titre produced by AP1-BP-AP2 fusion antigen was the highest, at up to 1:25600. The mice were then injected with 0.5 ml of 2 × 104 CFU/ml clinically isolated S. agalactiae at day 50 and observed daily for the following 7 days. RESULTS: Statistical analyses showed that these 4 kinds of fusion antigens had good protective immunity. Among them, AP1-BP-AP2 fusion antigen had the best protective immunity in Balb/c mice, with an immune protection index (PI) of 80%. Conclusion: This research provides a reliable theoretical basis for screening candidate antigens of the subunit vaccine and detecting antigen preparations of S. agalactiae.

Identification of genes and proteins associated with anagen wool growth.

Identifying genes of major effect for wool growth would offer strategies for improving the quality and increasing the yield of fine wool. In this study, we employed the Agilent Sheep Gene Expression Microarray and proteomic technology to investigate the gene expression patterns of body side skin (more wool growing) in Aohan fine wool sheep (a Chinese indigenous breed) in comparison with groin skin (no wool growing) at the anagen stage of the wool follicle. A microarray study revealed that 4772 probes were differentially expressed, including 2071 upregulated and 2701 downregulated probes, in the comparisons of body side skin vs. groin skin (S/G). The microarray results were verified by means of quantitative PCR. A total of 1099 probes were assigned to unique genes/transcripts. The number of distinct genes/transcripts (annotated) was 926, of which 352 were upregulated and 574 were downregulated. In S/G, 13 genes were upregulated by more than 10 fold, whereas 60 genes were downregulated by more than 10 fold. Further analysis revealed that the majority of the genes possibly related to the wool growth could be assigned to categories including regulation of cell division, intermediate filament, cytoskeletal part and growth factor activity. Several potential gene families may participate in hair growth regulation, including fibroblast growth factors, transforming growth factor-ß, WNTs, insulin-like growth factor, vascular endothelial growth factors and so on. Proteomic analysis also revealed 196 differentially expressed protein points, of which 121 were identified as single protein points.

Development of an indirect ELISA for bovine mastitis using Sip protein of Streptococcus agalactiae.

The sip gene encoding for a conserved highly immunogenic surface protein of Streptococcus agalactiae was amplified using polymerase chain reaction (PCR) and subcloned into prokaryotic expression vector pET32a (+) and expressed as a recombinant protein in E. coli BL21 (DE3). An indirect enzyme linked immunosorbent assay (ELISA) was developed using the purified Sip protein as a coating antigen, which could identify S. agalactiae specific antibody in sera. The coating antigen at a concentration of 3.125 µg/ml, serum diluted to 1:160, and HRP-conjugated secondary antibody concentration at 1:4000 was found to be most effective in exhibiting positive result. The ELISA was found to be highly specific for S. agalactiae that may be used for the detection of the pathogen in mastitis cases, for epidemiological studies and for surveillance.

ALK gene amplification is associated with poor prognosis in colorectal carcinoma.

BACKGROUND: Recently, the anaplastic lymphoma kinase (ALK) has been found to be altered in several solid and haematological tumours. ALK gene copy number changes and mutations in colorectal cancers (CRCs) are not well characterised. We aimed to study the prevalence of ALK copy number changes, translocations, gene mutations and protein expression in 770 CRC patients, and correlate these findings with molecular and clinico-pathological data. METHODS: ALK gene copy number variations and ALK expression were evaluated by fluorescence in situ hybridisation (FISH) and immunohistochemistry, respectively. RESULTS: Translocations of the ALK gene were not observed; 3.4% (26 out of 756) of the CRC patients tested had an increase in ALK gene copy number either amplification or gain. Interestingly, increased ALK gene copy number alteration was associated with poor prognosis (P=0.0135) and was an independent prognostic marker in multivariate Cox proportional hazards model. The study reveals a significant impact of ALK gene copy number alterations on the outcome of patients with CRC. CONCLUSION: The findings of our study highlight a potential role of targeting ALK in advanced CRCs by using ALK FISH and ALK IHC as a screening tool to detect ALK alterations. Based on these findings, a potential role of ALK inhibitor as a therapeutic agent in a subset of CRC merits further investigation.

Comparable efficacy of self-monitoring of quantitative urine glucose with self-monitoring of blood glucose on glycaemic control in non-insulin-treated type 2 diabetes.

AIM: To assess whether self-monitoring of quantitative urine glucose or blood glucose is effective, convenient and safe for glycaemic control in non-insulin treated type 2 diabetes. METHODS: Adults with non-insulin treated type 2 diabetes were recruited and randomized into three groups: Group A, self-monitoring with a quantitative urine glucose meter (n = 38); Group B, self monitoring with a blood glucose meter (n=35); Group C, the control group without self monitoring (n=35). All patients were followed up for six months, during which identical diabetes care was provided. RESULTS: There was a significant decrease in HbA1c within each group (p < 0.05). At the study conclusion, mean changes in HbA1c from baseline were -1.9% for Group A, -1.5% for Group B and -1.0% for Group C, and the proportion of patients achieving HbA1c ≤ 6.5% were 38.9%, 35.3% and 20.0% respectively. However, no significant differences between the groups were found. The average monitoring frequency was significantly higher in Group A than in Group B. The incidence of hypoglycaemia and quality of life scores were similar between the groups. CONCLUSIONS: This study suggests that self-monitoring of urine glucose has comparable efficacy on glycaemic control, and facilitates better compliance than blood self monitoring, without influencing the quality of life or risk of hypoglycaemia.

Correlation between cellular phone use and epithelial parotid gland malignancies.

The authors investigated the association between cellular phone use and epithelial parotid gland malignancy. The subjects were 136 cases who were treated for this condition at the authors' hospital from January 1993 to March 2010, and 2051 controls who did not have salivary gland tumours and were admitted to the oral and maxillofacial surgery department during the same period. Logistic analysis was used to examine the relationship between cellular phone use and risk of epithelial parotid gland malignancy and mucoepidermoid carcinoma. Overall, the frequency of cellular phone use was not significantly associated with epithelial parotid gland malignancy. Female gender, advanced age, married status, high income, and smoking were associated with an elevated risk of epithelial parotid gland malignancy, especially mucoepidermoid carcinoma. Residence in a rural area was associated with reduced risk of epithelial parotid gland malignancy. The results suggest a possible dose-response relationship of cellular phone use with epithelial parotid gland malignancy. The authors suggest that the association of cellular phone use and epithelial parotid gland malignancy and mucoepidermoid carcinoma requires further investigation with large prospective studies.

Role of circadian clocks in the development and therapeutics of cancer.

Circadian clocks are endogenous molecular time-keeping systems that underlie daily fluctuations in multiple physiological and biochemical processes. It is now well recognized that dysfunction of the circadian system may be associated with a heightened incidence of cancer. This brief review presents evidence supporting the important role played by circadian clocks in the development of cancer and the therapeutic efficacy of anticancer agents. A number of circadian clock genes have been identified, which include transcription factors that regulate gene expression. Continued research in this area should increase understanding of the role of circadian clocks which could, ultimately, reduce the incidence of cancer in people with disrupted sleep-wake cycles, such as 24-h shift workers and flight attendants, and provide optimal chronopharmacology for cancer treatment.

Genetic polymorphisms of MTHFR and aberrant promoter hypermethylation of the RASSF1A gene in bladder cancer risk in a Chinese population.

Epidemiological studies have shown that folate deficiency increases the risk of cancer by affecting DNA repair and methylation. Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in folate metabolism. In this study, it was hypothesized that MTHFR (C677T and A1298C) polymorphisms would be associated with bladder cancer and also with hypermethylation of the promoter of the Ras association domain family 1A (RASSF1A) gene. This hospital-based, case-control study of 312 bladder cancer patients and 325 cancer-free controls found that individuals carrying the MTHFR 677TT genotype had a 2.00-fold increased risk of bladder cancer compared with those carrying the 677CC genotype. None of the MTHFR A1298C polymorphisms alone were associated with bladder cancer, but the combined haplotype 677TT/1298AA was associated with a 2.27-fold increased risk compared with haplotype 677CC/1298AA. There was no association between MTHFR gene variants and methylation status of the RASSF1A gene in the 45 bladder cancer patients in whom this was studied. It is concluded that the MTHFR 677TT genotype and the TTAA haplotype may increase the risk of bladder cancer.

Styloid process osteochondroma.

Osteochondroma, a benign tumor of the skeleton and auxiliary tissues, is usually seen around the epiphysis of long bone, flat pelvis, scapula and transverse process of vertebra, but seldom in the head and neck region. The incidence of osteochondroma in the jaw bone only accounts for 0.6% of the whole body incidence, with rare reports of osteochondroma protruding into the oral cavity. Primary osteochondroma in the styloid process has not been reported until now. A case of styloid process osteochondroma and its surgical removal are here described.

Variable drug metabolism genes in Arab population.

Cataloging interethnic differences in the distribution of genotypes of drug metabolic genes provides valuable information for profiling the pharmacogenetics of a population. We used PCR analysis to catalog the frequencies of alleles and genotypes for CYP1A1, NAT2, GSTs, MTHFR, MTR (MS) and NQO*1 in Arabs. The frequencies of alleles and/or genotypes for CYP1A1*2A, GSTT1 null, GSTT1 and GSTM1 double null, and GSTP1 A1578G in Arabs were significantly higher than those reported in Caucasians. However, the distribution of NAT2 acetylator phenotypes in both populations was similar. In contrast, the frequencies of MTHFR 677T allele and the combined (677+1298) genotypes for low activity were lower than those reported in Caucasians. Other alleles in Arabs, including CYP1A1 T3801C and GSTP1 A1578G were present in frequencies similar to Africans. The overall profile of variations in metabolism genes in Arabs is thus unique.

[Study on the role of human papillomavirus in carcinogenesis of oral papillomas by in situ hybridization].

OBJECTIVE: To explore the role of human papillomavirus (HPV) infection on carcinogenesis of oral squamous cell papillomas (SCP). METHODS: By using digoxin labelled HPV 6/11 and HPV 16/18 probes, through hybridization in situ technique, the HPV DNA sequence in 30 cases of oral SCP were detected. RESULTS: Among them, 16 cases (53.3%) were positive for HPV 6/11 DNA. None of cases of SCP were positive for HPV 16/18 DNA. CONCLUSION: Our results show that the frequent finding of HPV 6/11 DNA in oral SCP had close association between oral papillomas and HPV 6/11 infection.

[A herbalogical study on traditional Mongolian medicine "lideri"].

By herbalogical study and investigation, "lideri" used by Mongolia doctors in different areas mainly contains 10 species from 4 genera of 4 families, but the quality materials only contains 3 species, Tinospora sinensis (Lour.) Merr., T. cordifolia Miers and T. capillipes Gagnep.

[A herbalogical study on traditional Mongolian medicine dakesha].

By herbological study and investigation, "Dekesha" used by Mongolia doctors mainly contains 17 species from 3 genus of 3 family, but "Dakesha" in Inner Mongolia Standard of Medicinal Materials only contains 2 species which are Oxytropis myriophylla and O. hirta.

[Quantitative 3D measurements of the skull base by using spiral volumatric CT images].

OBJECTIVE: To study the spatial relationships of mandibular condyle and carotid canal, jugular fossa, foramen ovale, foramen spinosum when mandible in normal physical position. METHODS: 100 native adults were examined by spiral volumatric CT. The data were transferred to workstation, three-dimensional(3d) cranial base images were reconstructed, 3D measurements were performed by using specific 3d CT landmarks. 12 points were defined and 28 distances and angles were measured in the skull base from submandibular viewpoint. The internal pole of condyle and the condylar long axis were regarded as the center point and 0 degree axis in the reference frame. RESULTS: Submandibular approach provided a clear vision of the internal pole of condyle and condylar long axis. The ECC was located at 12.01 +/- 2.71 mm, 5.14 +/- 1.22 degrees, Jug was located at 15.11 +/- 2.72 mm, -56.27 +/- 14.05 degrees according to the reference frame. CONCLUSIONS: Quantitative measurements of 3D skull base can be very precise. Mandibular approach is easy to show the mandibular condyle. The reference frame is helpful to located other important structures of skull base in operation.

[Changes in blood coagulation affected by rifampicin administration].

OBJECTIVE: To study the characteristics of influence on coagulation by rifampicin (RFP) administration. METHODS: Seventy-three patients, ages between 10 and 65 years(mean age 30.3 years), with tuberculosis of neck lymphnode were treated with 2HRZS/10HRE. The prothrombin time, activity, fibrinogen, II: C, V: C, VII: C, IX: C and alanine aminotransferase concentration in each case were observed during the period of RFP administration. RESULTS: The prolongation of the PT and the reduction of the PA, FIB concentration in 48 cases were observed after 5-30 days following RFP administration. These changes depended on drug dosage and duration, especially the severe influence on coagulation were detected when the RFP were reused after the ALT normalized in seventh to tenth months. The ALT was (18 +/- 12) U/L in the tenth month, but the PT was (19.2 +/- 3.9) s and the FIB was (1.8 +/- 0.5) g/L respectively. CONCLUSIONS: Not only the coagulation factors II, VII, IX, X, but also I V, VI and VIII were affected by RFP. The coagulation test is more sensitive than ALT test in the evaluation of liver injury. RFP should be stopped when FIB less than 1 g/L in spite of ALT in normal range.

Alterations in the level of phosphotyrosine signal transduction constituents in human parotid tumors.

Human parotid tumors were evaluated for the activation of the phosphotyrosine signaling pathway by Western blot, enzyme activity assay, and reverse transcriptase-polymerase chain reaction. Warthin's tumor and mucoepidermoid carcinomas had the greatest level of tyrosine phosphorylated proteins identified in plasma membrane fractions. These tumors, along with pleomorphic adenocarcinoma, showed high levels of membrane expression of the tyrosine kinase receptor, c-erbB-2, and phosphatidylinositol-3-kinase. Expression of the epidermal growth factor receptor was confined to normal tissue. The level of mRNA for c-erb was elevated only in mucoepidermoid carcinomas. Messenger RNA levels for ras were unchanged from control levels in all tumors, while the level of src mRNA was higher in the tumor samples than the normal parotid tissue. The activities of several signal transduction kinases, including protein kinase A and C were elevated in tumor tissue (7.7- to 18.9- and 0.4- to 3.7-fold higher, respectively), relative to surrounding normal tissue. While the level of glandular amylase was reduced (22%-0% of normal levels) in the tumor tissue, epidermal growth factor (EGF) and transforming growth factor-alpha (TGFalpha) content was dramatically higher in the neoplastic tissue (10- to 170-fold and 4.6- to 6.0-fold, respectively). These results suggest that with the presence of elevated levels of EGF, TGFalpha, and the oncoprotein receptor c-erbB-2 in the membrane of parotid tumors, cell proliferation and activation of the phosphotyrosine signal transduction pathway may involve autocrine stimulation through the expression of high levels of growth factor and receptor in the same tissue.

[Inhibitory effects of Na2SeO3 on Tca8113 cells in vitro and in vivo].

In the study, Na2SeO3 effectively limited the growth and proliferation of Tca8113 cells both in vitro and in vivo. The response was dependent on the dose, the starting administered time, exposed length of selenium and density of inoculated Tca8113 cells. At 1 microgram/ml dose of selenium, there were remarkable inhibitory effects while no detectable inhibitory effect to L929 cells. 1 microgram/ml dose for less than 24 hours, the growth and proliferative ability of Tca8113 cells was reversal whereas more than that period, was irreversal. In vivo experiment, the morbidity of transplanted tumors was remarkably depressed with Se-intraperitoneal injection. At 60 micrograms/ip dose, the weight of nude mice were not reduced and the pathological changes in liver and kidney had not found.

Clinical application of autogenous mandibular bone grafts. Analysis of 166 cases.

Of the 166 patients receiving autogenous mandibular bone grafts during 1972-1987, 55 were used for repair of mandibular bone defect, 67 for correction of maxillofacial malformation and 44 for reconstruction of temporo-mandibular joint. Follow-up showed that 155 patients had satisfactory results after operation.