Using chemically defined medium as the control, mechanism of corn steep liquor (CSL) in complex medium during glycerol production by Candida glycerinogenes was studied.The results showed that there were three key factors in CSL that had some great influences on glycerol fermentation of C.glycerinogenes, including phosphorus, nitrogen, and trace elements.The maximum glycerol yield of 53.44% was achieved at an optimal phosphorus concentration of 121.75mg/L, where the CSL concentration was 14g/L.Phosphorus in CSL could control the distribution of carbon metabolism flux between EMP pathway and HMP pathway.With the increase in CSL concentrations, superfluous phosphorus could restrain HMP pathway and activate EMP pathway, thus resulting in remarkable changes in various fermentation parameters of complex medium.Nitrogen in CSL could play a cooperative role in the regulative function of phosphorus.However, it was not a suitable nitrogen source for C.glycerinogenes.Trace elements in CSL could markedly improve the glucose consumption rate, accelerate the cell growth, and enhance the glycerol yield.

Animals ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; virology ; Myocardium ; metabolism ; Osteopontin ; metabolism ; Virus Diseases ; metabolism

OBJECTIVETo observe the effect of activation of aldehyde dehydrogenase 2 (ALDH2) by ethanol on the expression of c-Jun N-terminal kinase (JNK) in the kidney of diabetic rats.

METHODSEightheen healthy male SD rats were randomly divided into 3 groups (n = 6): normal control group, diabetes group and ethanol + diabetes group. After 8 weeks, 24 h urine samples from rats were collected to detect urinary protein content. The kidney was isolated and the ratio of kidney weight/body weight (index of kidney weight) was detected. The levels of fasting blood glucose, glycosylated hemoglobin serum urea nitrogen and serum creatinine were measured. Morphological changes of renal tissue were observed by optical microscope. The protein expressions of ALDH2 and JNK in renal tissue were detected by Western blot.

RESULTSCompared with the normal control rats, the levels of fasting blood glucose, glycosylated hemoglobin, serum urea nitrogen, serum creatinine and the index of kidney weight were increased markedly in diabetic rats. The expression of ALDH2 protein was decreased, while p-JNK, JNK protein expressions and the ratio of p-JNK/JNK were increased. The morphological observation was shown that the amount of glomerular mesangial matrix were increased, basement membrane were thickened and capillary lumen were narrowed. However,in ethanol + diabetes group, renal function was improved and the damage of renal structure was attenuated. The expression of ALDH2 protein was increased, while p-JNK, JNK and the ratio of p-JNK/JNK were decreased.

CONCLUSIONEnhanced ALDH2 expression can protect kidney in diabetic rats, which may be relevant with inhibitting the activity of JNK pathway.

Aldehyde Dehydrogenase ; metabolism ; physiology ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Diabetes Mellitus, Experimental ; enzymology ; Ethanol ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Kidney ; enzymology ; Male ; Mitochondrial Proteins ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

In MAST (mRNA accessible site tagging),the DNA tags from synthesized library were employed for identifying mRNA accessible sites. A large number of tags were amplified and subcloned for sequencing to verify mRNA binding profiles. A PCR was designed by using one primer which bridges over the tag terminal sequences. In PCR reaction DNA tag fragments were concatemerized by a bridge primer in reaction cycles. The concatemerized tag fragments were subcloned and sequenced. Dozens of the concatemerized sequences contained thousands tags. The PCR was a simple,effective way which for sequencing tags in a high through put manner.

Objective To evaluate the diagnostic value of urine Cystatin C(Cys C) for renal function impairment in neonates with hypoxic-ischemic encephalopathy(HIE).Methods The urine Cys C concentration was measured by enzyme linked immunosorbent assay(ELISA) in 47 cases of HIE newborns(25 cases were mild HIE and 22 cases were moderate-severe HIE) within 3 days after their birth.Twenty-three cases without perinatal asphyxia or other factors which could result in renal function impairment were selected as control group.Urine Cys C with urine retinal-bindingprotein(RBP),?2-microglobulin(?2-MG) and fractional sodium excretion(FENa%) were analyzed by kolmogorov-smirno in each group.Results Compared with control group,the concentration of urine Cys C,RBP and the levels of FENa% in HIE newborns were significantly elevated.The levels of urine Cys C in moderate-severe HIE newborns were significantly higher than those in mild HIE newborns(Pa

Psychrotrophs were isolated by using four media from low-temperature sewage of sewage treatment plant in Urumqi, Xinjiang. Totally, 154 strains were obtained including 12 filamentous fungi, 46 yeasts, 6 actinomycetes and 90 bacteria. The results of tolerance tests of the isolates to salt, phenol and SDS, and enzyme producing characters of amylase, proteinase and esterase were shown. Then 60 bacterial strains were chosen for 16S rRNA gene sequencing and analysis. The blasting results showed that the strains were assigned to 13 recognized genera , and the Strain 39 exhibited 96.6% similarity to Acinetobacter lwoffii(DSM2403), indicating that it might be a novel species. These results suggested that there were a lot of psychrotrophs and rich bacterial diversity in low-temperature sewage. In addition, which maybe an important and potential library of microbial resources.

Objective To investigate the effects of Plasmodium vivax merozoite surface protein 1(PvMSP1)on differentia-tion,maturation and function of dendritic cells(DC)and the mechanisms of PvMSP1 on the activation of DC via toll like receptors (TLR). Methods DCs were incubated with different doses of PvMSP1(1.0,10.0,100.0μg/ml)in vitro. The changes of CD83, CD86,and HLA-DR on DC were detected by flow cytometry(FCM);the expressions of cytokine IL-10 and IL-12 of DC were mea-sured by ELISA;the expressions of TLR4 and TLR9 mRNA of DC were measured by RT-PCR;the proliferation induction to autol-ogous lymphocytes of DC was measured by MTT. Meanwhile,the untreated DC and LPS inducing DC were as the negative control and positive control,respectively. All the data were analyzed statistically. Results Compared with the untreated DC,the propor-tions of CD83,CD86 and HLA-DR on DC induced by LPS and PvMSP1 increased significantly(all P<0.05);the expressions of IL-10 and IL-12 of DC induced by LPS increased significantly(P<0.01),and those induced by PvMSP1 also increased signifi-cantly(all P<0.05). In the LPS inducing group,the TLR4 mRNA production increased(P<0.05)and the TLR9 mRNA produc-tion had no significantly changes(P>0.05). In the PvMSP1-treated group,the DC TLR4 mRNA production increased(P<0.01) and the TLR9 mRNA production had no significantly changes(P>0.05);DC stimulated the proliferation of autologous lympho-cytes. Conclusion PvMSP1 enhances DC differentiation and maturation,and the mature DC induced by PvMSP1 has the ability of antigen presenting. The route for PvMSP1 inducing DC maturation might be TLR4 pathway rather than TLR9 pathway.

Eleven old male patients with hyperuricemia were collected ( hyperuricemia group,65-90 years old ).10 healthy middle-aged males ( middle-aged group,30-40 years old) and 10 healthy old males ( older group 60-70 years old ) with normal blood uric acid level were used as controls.All of the subjects were given low purine content diet ( 250 mg/d ) for 3 days followed by high purine content diet ( 800 mg/d ) consecutively for another three days.The samples of fasting blood and 24 h urine were collected for assay.The results showed that there were no significant changes of serum uric acid ( UA ) concentration in three groups after low purine content diet.But the levels of serum UA in three groups all increased significantly after high purine content diet,and the change was higher in hyperuricemia group than middle-aged group [ ( 507.7 ± 108.1 vs 378.9 ± 80.1 ) μmol/L,P＜0.05 ].24 h urine uric acid excretion in three groups was all significantly decreased after low purine content diet and increased after high purine content diet.After high purine content diet,24 h urine uric acid was lower in hyperuricemia group than middle-aged group [ ( 2.99 ± 1.21 vs 3.62 ± 1.02 ) mmol/24 h,P＜0.05 ].Blood urea nitrogen levels in all subjects decreased after low purine content diet and increased after high purine content diet ( P＜0.05 or P＜0.01 ).Creatinine clearance rate in hyperuricemia group was decreased after high purine content diet compared with baseline [ (75.3 ± 20.3 vs 80.7 ±20.0) ml/min ],and there were no significant changes in other groups after low and high purine content diet.24 h urine protein in hyperuricemia group was higher than middle-aged group ( P＜0.05 ),and increased after high purine content diet with significant difference ( P＜0.05 ).These results suggest that high purine content diet and decreased by renal uric acid clearance mainly contribute to hyperuricemia in old people.

Objective To construct an eukaryotic expression vector of retinoblastoma gene (pIRES-Rb94), and investigate the radiosensitising effect of Rb94 on lung adenocarcinoma cell line A549 and the mechanism. Methods Recombinant expression plasmid pIRES-Rb94 was constructed and then transfected into A549 cells using lipofectamine 2000. Steadily transfected cells were obtained using G418 se-lecting system. Cell counting method was used to produce the growth curve and the population doubling time was then calculated. The radiosensitivity of A549 cells was assessed by clonogenic assay. The expression of bTERT and Bcl-2 mRNA was detected by real-time RT-PCR. Cell cycle distribution was measured by flow cytometry. Results Steadily transfected cell line pIRES-Rb94-A549 was aquired. Compared with A549 cells, the population doubling time of pIRES-Rb94-A549 cells was increased from 31.5 h to 39.5 h (t=5.15, P<0.01). The expression of hTERT and Bcl-2 mRNA was both down-regulated (0.02:1.00, t= 18.99,P<0.01,0.01:1.00,t=13.73,P<0.01). The number of cells was increased in G2/M phases (35.91%:4.53%, t =36.78,P=0.00), whereas decreased in G0/G1 and S phases (47.02%:74.07%, t =11.71,P=0.00;17.07%:22.32%, t =2.30,P<0.05). The sensitizing enhancement ratio was 1.30. Conclusions Rb94 has marked radiosensitizing effect on A549 cells by G2/M phase blockage and down-regulation of hTERT and Bcl-2 mRNA expression. Rb94 may also inhibit the ability of cell proliferation by regulating cell cycle distribution.

OBJECTIVETo observe the effect of Chinese herbal extract HuNan A-1 (HNA-1) on the thymic output function in Simian immunodeficiency virus (SIV) chronically infected rhesus macaques.

METHODSEight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months, and then they were randomly divided into the treatment group and the control group, 4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage, once daily for 2 successive months, while those in the control group were administered with equal volume of normal saline by gastrogavage, once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts, as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles (TREC) were detected using real-time fluorescent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue, CD4 counts, naive CD counts, and TREC were analyzed.

RESULTSThere was no statistical difference in body weight, viral loads, absolute CD ratios between the two groups after treatment (P > 0.05). The altered TREC multiple showed an obvious decreasing tendency in the control group, while it showed an increasing tendency in the treatment group (P < 0.05). In both groups, destroyed structures of the thymus tissue could be seen, filled with pink unstructured material. Increased connective tissues, lowered connective cell density, and confused arrangement could also be seen in the two groups, with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum (r = 0.926, P = 0.001). Naive CD4 counts were positively correlated with CD4 counts (r = 0.961, P = 0.005).

CONCLUSIONSTREC content determination, as a marker of newly thymic emigrants, could be taken as a testing method for evaluating the thymic output function. Besides, HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents, naive CD4 counts, and pathologies of thymus tissues, especially in late infection stage.

Animals ; CD4 Lymphocyte Count ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Macaca mulatta ; Plant Extracts ; pharmacology ; Random Allocation ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; Simian Immunodeficiency Virus ; Thymus Gland ; drug effects ; Viral Load