Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.

In the basis of a large amount of literatures, this article sumed up the characteristics and application of eggshell membrane.

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

OBJECTIVETo observe the effects of blood activating wind dissipating acupuncture (BAWDA) on blood pressure (BP) of prehypertension (PHT) patients.

METHODSTotally 60 PHT patients were assigned to the control group and the acupuncture group according to random digit table, 30 in each group. All patients were intervened by life style. BAWDA was additionally performed in patients in the acupuncture group for 6 weeks (30 times). The improvement of BP after intervened by acupuncture was observed. BP success rates and the proportion of PHT progressing to hypertension (HT) were also observed after 6-week intervention of acupuncture and at 1-year follow-up.

RESULTSSystolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased after 6-week intervention in the acupuncture. The BP control rate was 56.7% (17/30 cases) in the acupuncture group vs.10.0% (3/30 cases) in the control group with statistical difference (chi2 = 14.70, P < 0.01). At 1-year follow-up BP success rate was 36.7% (11/30 cases) in the acupuncture group, remarkably higher than that of the control group [13.3%, (4/30 cases)] (chi2 = 4.36, P < 0.05).

CONCLUSIONSBAWDA showed BP regulating roles in a gradually stable decreasing tendency. It also could elevate BP success rate of PHT, and reduce the risk of PHT progressing to HT.

Acupuncture ; methods ; Acupuncture Therapy ; methods ; Blood Pressure ; Humans ; Hypertension ; Prehypertension ; therapy ; Wind

Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.

OBJECTIVETo investigate the antagonistic effects of three species of oral Streptococcus on the growth of oral Saccharomyces albicans in vitro.

METHODSDirect inoculation method, reverse inoculation method and mixed culture methods were respectively chosen to observe the changes of Saccharomyces albicans colony formation on the effects of Streptococcus mutans, Streptococcus sanguis and Streptococcus salivarius.

RESULTS1) No clear inhibition zone was observed in each of the groups by direct inoculation method. 2) Compared with the control groups, Saccharomyces albicans colony formation on soft agar of Streptococcus sanguis decreased significantly (P < 0.05). 3) Mixed culture method results showed that Streptococcus mutans could inhibit the growth of Saccharomyces albicans significantly at different time points (P = 0.001). 4) Under the action of bacteria culture supernatant, the count of Saccharomyces albicans in experiment groups showed statistical significance when compared with the control groups at 24, 48, 72 h (P = 0.001); The differences among the experimental groups were of no statistical significance at majority times (P > 0.05).

CONCLUSIONStreptococcus mutans, Streptococcus sanguis, and Streptococcus salivarius could obviously inhibit the growth of Saccharomyces albicans in vitro. However, it is still unclear that among which the inhibition effects is stronger. The antagonistic effects is weakened gradually.

In Vitro Techniques ; Saccharomyces ; Streptococcus ; Streptococcus mutans ; Streptococcus sanguis

OBJECTIVEThe aim of this study was to investigate whether the three species of oral Actinomyces have inhibitory effects on the growth of oral Candida albicans in vitro.

METHODSStraight o'clock method was used to observe the bacteriostasis circle. Reverse o'clock and mixed culture method were used to study the quantitative changes of Candida albicans colony respectively.

RESULTS(1) None of the groups had been viewed the bacteriostasis circle. (2) Compared with control groups, there was a significant decrease of Candida albicans colony on Actinomyces viscosus TPY soft agar (P < 0.05). Actinomyces naeslundii and Actinomyces odontolyticus TPY soft agar were both devoid of obvious Candida albicans colony (P < 0.01). The former group (Actinomyces viscosus) and the two latter groups (Actinomyces naeslundii and Actinomyces odontolyticus) showed a striking contrast (P < 0.01). (3) Compared with control groups, a decrease of Candida albicans showed up in the mixed culture, and the difference was significant (P < 0.05). The discrepancies among the three experimental groups were of no statistical value (P > 0.05).

CONCLUSIONOral Actinomyces viscosus, Actinomyces naeslundii and Actinomyces odontolyticus could inhibit the growth of Candida albicans in vitro. However, which of them contributed more to the inhibitory effects was still not affirmed.

Actinomyces ; Actinomyces viscosus ; Candida albicans ; In Vitro Techniques

OBJECTIVE To establish an ion-pair reversed-phase high performance liquid chromatography(HPLC) method and determine ATP,ADP and AMP in liver of the rats treated by electric acupuncture.METHODS Separation of ATP,ADP and AMP in the liver tissue samples of rats was performed on a Megres-C1 8 water resistant column(5 μm,250 mm×4.6 mm) by Shimadzu SIL-20AT.A mobile phase was 0.18 mol/L phosphate dihydrogen containing 5% methanol (pH =6.25).The detection wavelength was set at 254 nm and the flow rate was 0.8 mL/min.The injection volume was 20 μL and the column temperature was set at 25 ℃.RESULTS The linear ranges of ATP,ADP and AMP were 1.1~212.2 μg/mL(r =0.999 9), 0.9~180 μg/mL(r =0.999 9) and 0.9 ~ 181.8 μg/mL (r =0.999 7).The average recovery rates of ATP,ADP and AMP were 96.4%,98.3% and 100.8%;and RSD were 2.50%,2.88% and 4.14%,respectively.The RSD of intra-day and inter-day were 0.06%~0.40% and 0.06%~0.69%.The detection limits for ATP,ADP and AMP were 0.1 1,0.09 μg/mL and 0.045μg/mL,respectively.CONCLUSION The established method for the determination of ATP,ADP and AMP in the liver of rats is simple,quick,accurate and reliable,which will lay a foundation for the determination of small molecular bio-active substances in the animal tissues treated by acupuncture intervention.

BACKGROUNDResistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor.

METHODSThree cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide.

RESULTSThree cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation.

CONCLUSIONRBP could effectively rescue the promoted differentiation of resistin overexpressed 3T3-L1 preadipocyte.

3T3-L1 Cells ; Adipocytes ; cytology ; drug effects ; Amino Acid Sequence ; Animals ; Base Sequence ; Carrier Proteins ; isolation & purification ; pharmacology ; Cell Differentiation ; drug effects ; Mice ; Molecular Sequence Data ; Peptide Library ; Rats ; Resistin ; antagonists & inhibitors ; metabolism

BACKGROUNDTrichophyton rubrum (T. rubrum) represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that it may result from the failure of epithelial cells to recognize T. rubrum effectively and initiate effective immune responses. The C-type lectin receptors (CLR) and toll-like receptors (TLR) are the two major pattern recognition receptors (PRRs) that recognize fungal components. Therefore, the purpose of the study was to analyze the expression of those PRRs and the cytokines in HaCaT cells stimulated with heat-inactivated T. rubrum conidia and hyphae, respectively.

METHODSHaCaT cells were unstimulated or stimulated with heat-inactivated T. rubrum conidia and hyphae (1×10(6) and 1.5×10(5) colony-forming unit (CFU) in 2 ml medium, respectively) for 6, 12 and 24 hours. The mRNA expression of PRRs involved in recognizing fungal pathogen-associated molecular patterns (PAMPs) and signaling molecules were measured by quantitative reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, surface toll-like receptor (TLR) 2, TLR4 and Dectin-1 were analyzed by fluorescence-activated cell sorter (FACS) 24 hours after treatment. The cytokines were detected in cell culture supernatants of HaCaT cells in 12 and 24 hours after treatment.

RESULTSHaCaT cells constitutively expressed mRNA of membrane-bound TLR1, 2, 4 and 6, Dectin1 and DC-SIGN, but not Dectin-2 or Mincle. Heat-killed T. rubrum did not significantly upregulate gene transcriptions of the PRRs of HaCaT cells. Heat-inactivated T. rubrum conidia significantly reduced the surface expression of TLR2 and Dectin-1, and suppressed the secretions of interferon-inducible protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) of HaCaT cells, while heat-killed T. rubrum hyphae significantly induced the secretions of IP-10 and MCP-1.

CONCLUSIONThe cell-wall antigens of T. rubrum fail to activate transcriptional expression of PRRs and induce a lower immune response of HaCaT cells by limited cytokines secretion.

Cells, Cultured ; Cytokines ; biosynthesis ; Humans ; Keratinocytes ; immunology ; Lectins, C-Type ; genetics ; physiology ; RNA, Messenger ; analysis ; Receptors, Pattern Recognition ; genetics ; physiology ; Toll-Like Receptor 2 ; physiology ; Trichophyton ; immunology