The mechanisms of food allergy include IgE-mediated and non IgE-mediated immune responses.Food intolerance is the reaction of defected digestive function caused by many factors in which the immune system is not involved.Clinically,the cases of food intolerance are increasing and the symptoms of the disease are usually complicated.Clinicians are facing the problem of differential diagnosis of food allergy and food intolerance,this article reviews the mechanisms of these two diseases to provide the reference for differential diagnosis.

The phenylacetone monooxygenase, isolated from Thermobifida fusca, mainly catalyzes Baeyer-Villiger oxidation reaction towards aromatic compounds. Met446 plays a vital role in catalytic promiscuity, based on the structure and function of phenylacetone monooxygenase. Mutation in Met446 locus can offer enzyme new catalytic feature to activate C-H bond, oxidizing indole to finally generate indigo and indirubin, but the yield was only 1.89 mg/L. In order to further improve the biosynthesis efficiency of the whole-cell catalyst, metabolic engineering was applied to change glucose metabolism pathway of Escherichia coli. Blocking glucose isomerase gene pgi led to pentose phosphate pathway instead of the glycolytic pathway to become the major metabolic pathways of glucose, which provided more cofactor NADPH needed in enzymatic oxidation of indole. Engineering the host E. coli led to synthesis of indigo and indirubin efficiency further increased to 25 mg/L. Combination of protein and metabolic engineering to design efficient whole-cell catalysts not only improves the synthesis of indigo and indirubin, but also provides a novel strategy for whole-cell catalyst development.
Escherichia coli ; genetics ; metabolism ; Glucose ; metabolism ; Indigo Carmine ; metabolism ; Indoles ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; Metabolic Networks and Pathways ; Protein Engineering

OBJECTIVE This study was conducted to investigate the expression of Stat3 and its target gene products including Cyclin D1 and Bcl-xl in human laryngeal cancerous tissue, and to explore the mechanism in tumorigenesis of larymgeal carcinoma, and to explore the correlation between the expression,clinical parameters as well as the pathological parameters of laryngeal carcinoma. METHODS The expression of Stat3, p-Stat3, Cyclin D1 and Bcl- xl in 50 cases of cancerous tissues, adjacent normal tissues was measured by Western blot analysis. The expression pattern of Stat3 and its activated form p-Stat3 was determined by immunohisto- chemical staining. The correlation of the expression of Stat3, p-Stat3, Cyclin D1 and Bcl-xl in laryngeal carcinoma with various clinicopathological characteristics was analyzed statistically. RESULTS The protein expression level of stat3, p-Stat3, Cyclin D1, Bcl-xl (A value)were 86.97?1.58, 56.97?3.93, 24.43?5.62, 33.78? 3.56 in laryngeal carcinoma and 36.10?0.52, 16.52?0.52, 5.94?1.75, 14.68?2.14 in adjacent normal respectively(P

OBJECTIVE To construct Hep-2 cell line with stable transfection of siRNA-STAT3 gene, and to explore the relationship between STAT3 expressionand the pathogenesis of laryngeal cancer. METHODS Specific STAT3 oligonucleotides were designed and synthesized. These oligonucleotides were annealed form the double strands DNA fragments. Then the fragments were cloned into pGPU6/GFP/Neo vector. The recombinant PGPU6/GFP/Neo-siRNA-STAT3 plasmid was confirmed by enzyme digestion and sequence analysis at the same time. Positive clones were selected out by G418 and p-STAT3 expression was identified by Western-blot analysis. The growth curve of the Hep-2 cells was drawn based on MTT assay and the growth ability of single Hep-2 cell was measured according to the plate colony formation assay respectively. RESULTS PGPU6/GFP/NEO-siRNA-STAT3 expression vector was successfully constructed. Western-blot analysis identified that p-STAT3 expression declined obviously in siRNA- STAT3 group compared with that in negative control and blank group. The growth curve explained that the Hep-2 cells of siRNA-STAT3 group began to show obvious inhibitory effect until they were inoculated in the culture plate for 3 days(P =0.001).There were pronounced inhibitory effect after 5 days(P=0.000). The results also showed time-effect relationship of the inhibition. Furthermore, the cell colony formation rate of siRNA- STAT3 group was less than the negative control and blank group(P =0.000). CONCLUSION We successfully selected out Hep-2 cell line expressed siRNA-STAT3 gene stably and efficiently in this study. Down regulation of STAT3 correlated with the inhibition of growth of Hep-2 cells.

Purpose: To investigate the clinical effect of Yin-reinforcing and Blood-nourishing Drink plus electroacupuncture in the treatment of senile skin pruritus. Methods: Eighty patients were randomly divided into two groups. The treatment group was treated with Yin-reinforcing and Blood-nourishing Drink plus electroacupuncture and the control group, with Western medicine. Results: The total effective rate was 100.0% in the treatment group and 67.5% in the control group (P＜0.01). Conclusion: This therapy has a better effect on senile skin pruritus.

Objective To com pare and explore the application value of diatom nitric acid digestion method and plankton 16S rDNA PCR method for drow ning identification. Methods Forty drow ning cases from 2010 to 2011 were collected from Department of Forensic Medicine of Wenzhou Medical University. Sam ples including lung, kidney, liver and field water fromeach case were tested with diatom nitric acid digestion method and plankton 16S rDNAPCR method, respectively. The Diatom nitric acid digestion method and plankton 16S rDNAPCR method required 20 gand 2g of each organ,and 15 mL and 1.5 mL of field water, respectively. The inspection time and detection rate were com pared between the two methods. Results Diatom nitric acid digestion method m ainly detected two species of diatom s, Centriae and Pennatae, while plankton 16S rDNA PCR method am plified a length of 162 bp band. The average inspection time of each case of the Diatom nitric acid digestion method was (95.30±2.78) min less than (325.33±14.18)min of plankton 16S rDNA PCR method (P<0.05).The detection rates of two methods for field water and lung were both 100% . For liver and kidney, the detection rate of plankton 16S rDNA PCR method was both 80% , higher than 40% and 30% of diatom nitric acid digestion method (P<0.05), respectively. Conclusion The laboratory testing method needs to be appropriately selected according to the specific circum stances in the forensic appraisal of drow ning. Com pared with diatom nitric acid digestion method, plankton 16S rDNA PCR method has practice values with such advantages as less quantity of sam ples, huge inform ation and high specificity.

Objective To evaluate the efficacy of double filtration plasmapheresis (DFPP) in the treatment of patients with refractory rheumatoid arthritis (RA). Methods Eighty-two patients were randomly aesigned,42 to the DFPP group and 40 to the no-DFPP group. All patients previously experienced an incomplete response to 2-3 dis-ease-modifying antirheumatic drugs (DMARDs) and 1-2 nonsteroidal anti-inflammatory drugs (NSAIDs) or predni-sene. All patients received sulphasalazine (SASP,0.75 g three times daily) plus methotrexate (MTX, 10 mg orally once weekly). DFPP was performed once a week for 2-3 sessions. A total of 121 plasmapheresis procedures were per-formed in 42 patients. Control patients did not receive sham DFPP. The efficacy measures recorded one day after the final treatment and latest month in follow up for 12～24 months included the American College of Rheumatology 20% ,50% ,and 70% improvement criteria (ACR20, ACR50, and ACR70), the Health Assessment Questionnaire estimate of disability (HAQ); and the disease activity index. Results Patients in the DFPP group had ACR 20, ACR 50 and ACR 70 improvements of 100% ,92.9% and 81.0%,as compared with the patients in no-DFPP group 17.5% ,0,and 0 (P<0.001). Significant change from baseline was observed in HAQ scores in the DFPP group but not in the no-DFPP group (P<0.001). The changes from baseline in the disease activity scores were significantlygreater than in the no-DFPP group (P<0.001). Conclusion DFPP therapy significantly alters the signs and symp-toms of refractory RA. There are significant increases in physical function and improvement in quality of life.

OBJECTIVE: To show that Stat3 played a key role in the G1 to S phase transition in laryngocarcinoma cells. METHOD: Human laryngocarcinoma cell lines Hep-2 were transfected with Stat3 antisense oligonucleotide mediated by liposome, MTT assay was used to measure the proliferation, flow cytometry was applied to analyze the cell cycle, and the expressions of Stat3, phosphorylation specific Stat3 (tyrosine705), CyclinD1, Cyclin E, CDK2, CDK4, CDK6, p21 and p27 were detected by western blot. RESULT: Hep-2 laryngocarcinoma cell lines expressed constitutively activated Stat3. Antisense oligonucleotide which directed blocked up the translation site resulted in growth inhibition, downregulation of Stat3, p-Stat3, Cyclins and CDKs, and upregulation of p21 and p27. CONCLUSION Our findings suggested that Stat3 played an important role in the G1 to S phase transition in laryngocarcinoma cells, Stat3 orchestrated cell cycle by regulating the balance between CDK/Cyclin complex and CKI.
Cell Line, Tumor ; G1 Phase ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; S Phase ; STAT3 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; Transfection

Objective To estimate the value of detection of Merkel cell carcinoma polyomavirus (MCPyV)in the diagnosis of Merkel cell carcinoma (MCC).Methods Two cases of MCC were studied using light microscopy and immunohistochemistry.PCR was performed to detect DNA sequences encoding MCPyV large T antigen(LT)and viral protein 1 (VP1) in paraffin-embedded tissue specimens from the two patients with MCC,five patients with T cell lymphoma,two normal human controls,as well as in two T cell lymphoma cell lines MAC 1 and MAC2.DNA sequencing was also carried out.Results Both of the patients with MCC were male.The patient 1 presented with a mass in the right anterior shin for more than one year,and the patient 2 had a mass in the left knee for more than six months.Skin examination revealed densely distributed pink nodules in the right anterior shin,with confluence into an indurated plaque which measured 10 cm × 8 cm with superficial erosion,exudates and crusts and was surrounded by multiple irregularly sized erythematous nodules with limited mobility in the patient 1,as well as a royal blue,hard,poorly marginated nodular mass measuring 5 cm × 4 cm in the left medial knee with limited mobility in the patient 2.Pathological manifestations were similar in the two patients.Tumor cells were uniform with large hyperchromatic nuclei,eosinophilic and sparse cytoplasm,and fine chromatin.Mitotic figures were easily seen.Immunohistochemistry revealed that the tumor cells stained positive for pan-cytokeratin,synuclein (Syn),neuron-specific enolase (NSE),chromogranin (CgA),CK20,and Ki-67 (＞ or =60％),but negative for S100 protein,HMB45,CD34,thyroid transcription factor 1 (TTT-1),CK7 and leukocyte common antigen (LCA).MCPyV DNA was detected in both MCC specimens,but absent in the other skin specimens or T cell lymphoma cell lines.Conclusions MCC has distinctive clinical and pathological appearance.Immunohistochemistry and detection of MCPyV DNA sequences using PCR may be beneficial to the definitive diagnosis of MCC.

[ABSTRACT]OBJECTIVETo investigate the relationship between food allergen specific IgE, IgG and allergic rhinitis, and to explore the significance of food allergen in the diagnosis and prevention of allergic rhinitis.METHODSTen kinds of food specific IgE were detected by Western blot and 14 kinds of food specific IgG were detected by ELISA in 2860 allergic rhinitis patients. The positive results of specific IgE and specific IgG were retrospectively analyzed.RESULTSThere was no difference in positive rate of allergen in patients with different gender. Total positive rate of food specific IgE was 68.5%. The specific IgE positive rate of crab was 31.5%, shrimp 27.6%, milk 21.3%, fish assemblages 19.2%, freshwater fish assemblages 18.3% and soybean 17.2%. With the growth of age, the specific IgE positive rate of peanut increased, but eggs decreased. The total positive rate of food allergen specific IgG was 81.5%. The specific IgG positive rate of egg was 59.4%, milk 38.2%, codfish 32.6%, soybean 22.8%, rice 19% and shrimp 12.7%. With the growth of age, the specific IgG positive rate of crab increased. Specific IgE and IgG of soybean, milk, crab, shrimp and fish were correlated well.CONCLUSIONThe detection of specific food allergen antibodies is helpful for the diagnosis of allergic rhinitis.