In industrial glutamate fermentation, intermitted feeding glucose with the help of off-line glucose measurement is generally necessary. This kind of feeding strategy could cause large variations in glucose concentration so that it is not favorable for the achievement of efficient and stable glutamate fermentation. Glutamate fermentation is characterized with typical non-growth association behavior, and during glutamate production phase glucose consumption is closely correlated with ammonia consumption. In this study, glucose concentration was controlled at various pre-determined levels by predicting glucose consumption amount and thus its concentration with the aid of on-line monitoring ammonia consumption. When glucose concentration was controlled around a lower level of 5 g/L~10 g/L, the final glutamate concentration could reach a relatively higher level of 80 g/L. In this way, the huge osmotic stress change due to the large glucose concentration variation with the intermitted feeding method could be avoided and the glutamate fermentation performance enhancement be expected.

OBJECTIVETo analyze the dental care utilization and expenditure of residents in Beijing, and to provide some basis on the policy of oral health insurance system.

METHODSA cross-sectional survey was conducted among 1,517 subjects (urban area) and 1,878 subjects (rural area) of all age groups in Beijing selected by stratified, clustering, random sampling. The data of oral health care utilization and expenditure were collected in their home.

RESULTSThe number of the people who visited a dentist in a year were low both in urban area and in rural area, but the expenditure for oral health care per visit were quite high. The value of utilization of dental care in rural residents was 1/3 of that in urban residents, while the value of expenditure in rural people was about 1/2 of that in urban people. 2.07% incomes of rural residents were used for dental care per year, the corresponding value of urban residents was 1.77%. There was significant difference on the expenditure among those with different demographic, socio-economic backgrounds.

CONCLUSIONSThe expenditure for oral health care was high in Beijing, which accounted for quite a lot in average incomes per year. The burden of expenditure for dental care on rural residents was heavier than that on urban residents. The level of expenditure for dental care could provide some references for oral health insurance system in Beijing.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Dental Health Services ; economics ; statistics & numerical data ; Fees, Dental ; Female ; Health Services Accessibility ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Insurance, Dental ; Male ; Middle Aged ; Rural Health Services ; statistics & numerical data ; Urban Health Services ; statistics & numerical data

To seek the reason of heterogeneity of recombinant HWTX-I (rHWTX-I) expressed in Pichia pastoris. We expressed HWTX-I gene of interest in Pichia pastoris GS115/HWTX-I. The heterogenous product expressed was separated, purified and identified by using Ion exchange HPLC, reverse HPLC, Tricine SDS-PAGE and MALDI-TOF Mass Spectrometry and then sequenced in both N-terminus and C-terminus. These results show that the heterogeneity of rHWTX-I results from the incomplete processing of signal peptide of N-terminus and the internal degradation of C-terminus. Biological activity assay shows that the activity of the heterogenous rHWTX-I only showed 30% activity compared with the native HWTX-I. The Solutions to how to avoid the heterogeneity are also discussed.
Animals ; Neurotoxins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Reptilian Proteins ; biosynthesis ; genetics ; Spider Venoms ; biosynthesis ; genetics

OBJECTIVETo prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity.

METHODSRecombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay.

RESULTSELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%.

CONCLUSIONWe obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.

Animals ; Antibodies ; immunology ; Antibody Formation ; Egg Proteins ; immunology ; Female ; Immune Sera ; Male ; Membrane Glycoproteins ; immunology ; Mice ; Oocytes ; Rabbits ; Receptors, Cell Surface ; immunology ; Recombinant Proteins ; immunology ; Sperm-Ovum Interactions ; Spermatozoa ; Zona Pellucida Glycoproteins

OBJECTIVETo investigate the relationship between the levels of antidesmoglein (DSG) 1, 3 antibodies in the sera of patients with paraneoplastic pemphigus (PNP) and alopecia.

METHODSSera from PNP patients, bullous pemphigoid patients, and normal healthy subjects were collected and 2 tissue samples from 2 healthy scalps were resected. Anti-DSG 1, 3 antibodies in the sera of PNP patients were detected by enzyme-linked immunosorbent assay (ELISA). Indirect immunofluorescent assay was used to detect whether the antibodies in the sera of PNP patients binds with the follicular epithelium of normal healthy scalp.

RESULTSAnti-DSG3 autoantibody was strongly positive and anti-DSG1 weakly positive in one patient, while both two antibodies were negative in the other patient. Their sera could bind to keratinocytes and follicular epithelium in human scalp. Immunofluorescent signals were found on the intercellular epidermal cell surface and outer root sheath of the follicular epithelium. However, the immunofluorescent signals in the section incubating with serum of bullous pemphigoid were only found on basal membrane zone. No signals were found in the section incubating with normal healthy serum.

CONCLUSIONAlopecia in PNP patients are correlated with the anti-DSG3.

Adult ; Alopecia ; etiology ; immunology ; Autoantibodies ; blood ; Desmoglein 1 ; immunology ; Desmoglein 3 ; immunology ; Female ; Humans ; Male ; Paraneoplastic Syndromes ; complications ; immunology ; Pemphigus ; complications ; immunology

The aim of this study was to investigate the effects of TLR2, TLR9, CD4(+)CD25(+) regulatory T cells (Treg) and transcription factor FoxP3 in the pathogenesis of children with infectious mononucleosis (IM). Thirty-five acute IM patients admitted in our hospital from April 2010 to January 2011 were enrolled in this study. Thirty-five healthy subjects were taken as control. The thirty-five patients before treatment were considered as patients in acute stage, after treatment and without clinical symptom they were thought as patients in recovery stage. The expression levels of TLR2, TLR9 and FoxP3 mRNA were detected by real time PCR using SYBR Green I. The expression of T lymphocyte subset CD4(+)CD25(+) in peripheral blood mononuclear cells was detected by flow cytometry. The results showed that the relative levels of TLR2 mRNA (4.03 ± 0.56), TLR9 mRNA (8.88 ± 1.56) in peripheral blood mononuclear cells of IM patients in acute stage were significantly higher than those of the controls [TLR2 mRNA (2.22 ± 0.57), TLR9 mRNA (3.63 ± 1.30)] and IM patients in recovery stage [TLR2 mRNA (2.76 ± 0.83), TLR9 mRNA (5.34 ± 1.60)] (P < 0.01). The result of CD4(+)CD25(+) (2.38 ± 1.32%) and relative level of FoxP3 mRNA(2.82 ± 0.90) in peripheral blood mononuclear cells of IM patients in acute stage were lower than those of the control [CD4(+)CD25(+) (7.85 ± 1.97%), FoxP3 mRNA (4.65 ± 1.23) ] (P < 0.01). There was no significant difference in CD4(+)CD25(+) (6.81 ± 1.84%), FoxP3 mRNA(4.11 ± 1.37) levels between IM patients in recovery stage and the controls (P > 0.05). It is concluded that the expression of CD4(+)CD25(+)regulatory T cells is reduced, and its special transcription factor FoxP3 mRNA is down-regulated, but expression levels of TLR2 mRNA, TLR9 mRNA are up-regulated in IM patients of acute stage.
Case-Control Studies ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Forkhead Transcription Factors ; metabolism ; Humans ; Infectious Mononucleosis ; diagnosis ; immunology ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Male ; RNA, Messenger ; metabolism ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 9 ; metabolism

Objective:Utilizing multi-criterion optimization (MCO) technology to improve plan design quality based on knowledge-based planning (KBP) model.Methods:Fifty-five patients with nasopharyngeal carcinoma (NPC) who had completed radiotherapy were selected, and fixed-field intensity-modulated radiotherapy (IMRT) technology was used in each case. Among them, 40 cases were randomly selected as training set 1. Then, IMRT plans in training set 1 were preprocessed by MCO technology to construct a new training set 2. With the initial training set 1 and the processed training set 2 as training samples, the traditional KBP model and the MCO-KBP model refined by MCO technology were trained, respectively. Among the remaining 15 cases, 5 cases were randomly selected as the validation set, and the remaining 10 cases were used as the test set. After verification, the test set was used to statistically analyze the plan quality of the initial manual plan and the automatic plan generated by the traditional KBP model and the MCO-KBP model.Results:The target dose (D 95%) of plans generated by the traditional KBP model and the MCO-KBP model met the clinical requirements. Conformity index (CI) and homogeneity index (HI) were almost the same ( P>0.05), and the doses of organ at risk (OAR) of the automatic plans generated by the MCO-KBP model were lower than those of the traditional KBP model. For example, compared with the traditional KBP model, the average D max of the brainstem in the automatic plans generated by the MCO-KBP model was lower by 2.13 Gy, the average D mean of the left parotid gland was lower by 1.39 Gy, the average D mean of the right parotid gland was lower by 1.59 Gy, and the average D max of the left optic nerve was lower by 1.42 Gy, the average D max of the right optic nerve was lower by 1.16 Gy, and the average D max of the pituitary gland was lower by 1.88 Gy. All of the above-mentioned dosimetry indexes were statistically significant. Conclusion:Compared with the traditional KBP model, the IMRT plans designed by the refined MCO-KBP model have obvious advantages in the protection of OAR, which proves the feasibility of utilizing MCO technology to improve the plan design quality of the KBP model.

OBJECTIVETo describe the distribution of cerebral vascular hemodynamic indexes (CVHI).

METHODSA number of 25,355 age 35 and over were selected in the Northeast China by cluster sampling. CVHI were checked during baseline survey and were followed to see the occurrence of stroke. Distribution of CVHI among non-stroke population, individuals prior to the onset of stroke and patients with stroke were described.

RESULTSThe CVHI accumulative score, V(mean), V(max) and V(min) were dramatically decreasing, but RV, Zcv, WV and DR were significantly increasing as age increased. V(max), RV and CP were significantly higher in males but WV was lower than that of females. The CVHI accumulative score, V(min) and RV were 95.0, 10.23 and 75.8 in non-stroke population, 51.25, 6.71 and 122.72 pre stroke group, and 55.0, 6.78 and 115.89 in patients with stroke respectively. There were significant differences among three groups after controlling of age and sex (P < 0.01).

CONCLUSIONVariance of CVHI was closely related to age, and there appeared a significant abnormal of CVHI before and after stroke.

Age Factors ; Aged ; Cluster Analysis ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Risk Factors ; Sex Factors ; Stroke ; physiopathology

OBJECTIVETo determine the effects of Ginkgo biloba extract (EGB) on major periodontal pathogens in subgingival plaque.

METHODSSixty patients with moderate to severe periodontitis were selected and randomly assigned to 3 groups: EGB group, a positive (periocline) and a negative control groups. Subgingival plaque samples were collected before treatment and 1 week, 2 months and 4 months after treatment. The detection rates of 4 major periodontal pathogens-Treponema denticola (Td), Tannerella forsythus (Tf), Prevotella intermedia (Pi), and Porphyromonas gingivalis (Pg)-were detected by polymerase chain reaction (PCR). Clinical indicators were examined before treatment, 3 and 6 months after treatment.

RESULTSEGB significantly decreased the detection rate of all the 4 pathogens 1 week after treatment, and then gradually increased at 2 and 4 months. EGB's inhibition effect was better than or comparable to periocline, except for Pg in short-term. The difference of plaque index (PLI) and bleeding index (BI) was not statistically significant among the groups, while for probing depth (PD) and attachment loss (AL), the difference was statistically significant between the EGB group and negative control group at 3 and 6 months after treatment.

CONCLUSIONEGB significantly inhibited major periodontal pathogens and can be used as an adjuvant for periodontitis treatment.

Adjuvants, Pharmaceutic ; pharmacology ; therapeutic use ; Bacteria ; drug effects ; isolation & purification ; Dental Plaque ; drug therapy ; microbiology ; Follow-Up Studies ; Ginkgo biloba ; chemistry ; Humans ; Periodontium ; drug effects ; microbiology ; pathology ; Plant Extracts ; pharmacology ; therapeutic use ; Polymerase Chain Reaction ; Treatment Outcome

OBJECTIVETo explore the feasibility of application of adipose-derived cells (ADCs) in reconstruction of tissue engineered cartilage in vitro.

METHODSAdipose tissue were obtained from human liposuction aspirate (19 cases, 31.5 +/- 5.8 years old). ADCs were isolated by collagenase digestion from liposuction aspirates. 3rd passage cells were seeded into PLGA scaffolds. The copolymer constructs were cultured in conditioned or non-conditioned medium in vitro for 4 weeks. The constructs were evaluated though gross morphology, histology, and immunohistochemistry.

RESULTSThe cell-polymer constructs kept its original shape in the induced group, but lost its original shape in the non-induced group. The scaffold group were collapsed. Histologically, the induced groups showed dense cellularity and lacunae-containing cells embedded in a basophilic matrix, while non-induced groups showed connective tissue-like morphology. Collagen and proteoglycan deposition was revealed by Massons's trichome and Safranin' O staining, and minor collagen II expression in the matrix was detected by immunohistochemistry staining in the induced group. They were all negative in the non-induced groups.

CONCLUSIONSAlthough ADCs included many kinds of cells, it is feasible to use ADCs as seeds cells for reconstruction of tissue engineered cartilage.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Adult ; Cartilage ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Female ; Humans ; Stromal Cells ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds