The 11-hydroxylation of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione as a useful intermediate for the preparation of hormones can be achieved by the mycelium of Absidia coerulea at higher conversion rate than using other strains. In this paper 16alpha,17alpha-epoxy-4-pregenene-3,20-dione mixed with a little water, beta-cyclodextrin, Tween-80 was introduced into the fermentation broth after ultrasonication to increase pseudo-water-solubility of the hydrophobic substrate. This pseudo-crystallo feed could avoid the toxicity of organic solvents and was more available for the microbial transformation. The multi layer feed-forward neural network was used to setup a model which indicated the relationship between medium and feed components and the conversion rate. Particle swarm optimization (PSO), which was a stochastic global optimization algorithm and of which the convergence speed was high, was applied to obtain the optimal concentration of the medium and feed components. At optimum conditions with the pseudo-crystallo feed, the conversion rate of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione at an initial concentration of 10 g/L was 87.5% in shaking flasks. The conversion rate of the substrate was up to 86.6% at higher concentration of 20 g/L feed in a 3.7 L fermentor.
Absidia ; metabolism ; Fermentation ; Hydroxylation ; Pregnenediones ; metabolism

Objective To explore the risk factors of skeletal related events (SREs) in non small cell lung cancer with bone metastases and its effect on the prognosis .Methods Totally 223 cases of NSCLC patients with bone metastasis were retrospective studied from January 2010 to December 2012 in our hospital .The clinical features ,predictive factors for SREs were analysed by sin‐gle factor and multifactor analysis .Results Among 223 cases of NSCLC patients with bone metastasis ,119 cases occured with SREs(53 .4% ) .Univariate analysis showed that the occurrence of SREs in female ,no smoker ,adenocarcinoma ,solitary bone metas‐tasis lesions were less than the male ,smoker non‐adenocarcinoma ,and multiple bone metastases (P<0 .05) ,but the rost without statistically significant(P>0 .05) .The multivariate analysis revealed only multiple bone metastases was an independent risk factor for SREs .The median survival time of the NSCLC patients with bone metastasis was 15 .3 months .Moreover ,survival analysis showed that SREs had no statistical significance on the prognosis of bone metastasis in NSCLC patients (P>0 .05) .Conclusion The female ,adenocarcinoma ,smoking history ,solitary bone metastasis lesions occurred in patients with lower risk SREs .Multiple bone metastasis is an independent risk factor for SREs ,attention should be paid to monitoring and prevention .

Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.

OBJECTIVETo determine the expressions of miR-155, miR-34a and miR-30a in diffuse large B-cell lymphoma (DLBCL) and to explore their potential correlation with clinicopathological characteristics.

METHODSThe expression level of miR-155, miR-34a and miR-30a in 46 DLBCL samples were determined with TaqMan real-time polymerase chain reaction. Interphase fluorescence in situ hybridization (I-FISH) was performed to detect MYC and p53 genes' status, and immunohistochemistry (Envision method) was used to evaluate the expression of CD3, CD10, CD20, BCL-6 and MUM-1 in DLBCL. The DLBCLs were classified into germinal center B cell-like (GCB) and non germinal center B cell-like (non-GCB) subtypes according to Hans' criteria.

RESULTSCompared with normal controls, miR-155 expression level was significantly higher in DLBCL. The expression level of miR-155 in non-GCB type was higher than that in GCB type. It was shown that the patients with MYC rearrangement had lower expression level of miR-155 than the negative controls. Compared with p53 normal group, the expression level of miR-34a was significantly lower in p53 deletion group. It was also shown that the patients with BCL-6 protein expression had lower expression of miR-30a compared with the negative group.

CONCLUSIONmiR-155 expression level is different in normal controls, DLBCL and patients with subtype DLBCL. It therefore has a diagnosis value for DLBCL. miR-34a is of great prognostic significance. miR-155, miR-34a and miR-30a may be potential therapy targets for DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

OBJECTIVETo analyze the role of resistin in insulin resistance (IR) through investigating the variation of plasma resistin levels and single-nucleotide polymorphisms (SNPs) in resistin gene 5' flanking region in stroke patients.

METHODSIn 103 atherothrombotic cerebral infarction (ACI) patients, 85 lacunar infarction (LI) patients, 70 intracerebral hemorrhage (ICH) patients, and 86 healthy controls, plasma resistin and insulin levels were measured by ELISA, SNPs in resistin gene 5' flanking region were detected by PCR and direct DNA sequencing. The subjects' body height and weight, the body mass index, quantitative insulin sensitivity check index (QUICKI), blood pressure, and the concentration of fasting plasma glucose, triglyceride, total cholesterol, creatinine, low-density lipoprotein, and high-density lipoprotein were also determined.

RESULTSQUICKI was significantly lower in the ACI and ICH patients (0.316 +/- 0.037 and 0.309 +/- 0.032, respectively) than that in the controls (0.342 +/- 0.043, P < 0.001), while plasma resistin level was significantly higher in the ACI and ICH patients (6.36 +/- 3.79 and 7.15 +/- 4.27 ng/mL, respectively) than that in the controls (5.28 +/- 2.56 ng/mL, P < 0.05), but such difference was not observed in the LI patients compared with controls. There was a statistically negative correlation between plasma resistin level with QUICKI (r = -0.228, P < 0.001). The distributions of allele and genotype frequencies of resistin gene - 420C > G and - 537A > C SNPs were not significantly different among the different groups, and those SNPs were not correlated with other clinical and biochemical parameters.

CONCLUSIONSPlasma resistin is associated with stroke by participating in the development of IR. The SNPs in resistin gene 5' flanking region has no impact on the plasma resistin level.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; metabolism ; Cerebral Infarction ; blood ; genetics ; Creatinine ; blood ; Female ; Humans ; Insulin ; blood ; Insulin Resistance ; physiology ; Intracranial Arteriosclerosis ; blood ; genetics ; Lipoproteins ; blood ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Resistin ; blood ; genetics ; Stroke ; blood ; genetics ; Triglycerides ; blood

OBJECTIVETo investigate BCL-6, MYC and p53 genes abnormalities in diffuse large B-cell lymphoma (DLBCL) and correlate the result with immunosubtypes and prognosis.

METHODSInterphase fluorescence in situ hybridization (I-FISH) was performed to detect the BCL-6, MYC and p53 genes. Immunohistochemistry (Envision method) was used to measure the expressions of CD3, CD10, CD20, BCL-6, MUM -1, BCL-2 and Ki-67 genes in DLBCL. The patients were classified into germinal center B cell-like (GCB) and non-GCB subtypes according to Hans' algorithm.

RESULTSBCL-6 rearrangement was detected in 10 of 46 DLBCL cases. The presence of gene rearrangement had no correlation with BCL-6 protein expression (P= 0.245). Overall survival (OS, P= 0.138) and progression-free survival (PFS, P= 0.095) were not influenced by BCL-6 rearrangement. All MYC rearrangements were detected in GCB type DLBCL. Deletion of p53 gene was detected in 14 cases and was significantly associated with shorter OS (P= 0.046) and PFS (P= 0.043).

CONCLUSIONI-FISH is a rapid, accurate and sensitive method for detecting BCL-6, MYC and p53 abnormalities. No correlation was found between BCL-6 gene rearrangement and BCL-6 protein expression. MYC translocation was more common in GCB type DLBCL compared with non-GCB type ones. Patients with p53 deletion had a poorer prognosis. The p53 gene may provide a useful indicator for the prognosis of DLBCL.

DNA-Binding Proteins ; genetics ; Female ; Genes, p53 ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; classification ; genetics ; immunology ; mortality ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Proto-Oncogene Proteins c-myc ; genetics

Objective To explore the relationship between -262C/T and -21A/T polymorphisms of catalase(CAT) gene and coal-burning borne fluorosis. Methods In 2007, 150 villagers were taken as a nonintervention group in Bijie city from the village of coal-burning borne fluorosis areas with unchanged cooking stoves;150 villagers were taken as the intervention group from the town of Changchun county where cooking stoves changed; 150 villagers were taken as control from non-endemic fluorosis areas in Baiyun town of Changshun county.PCR-restriction fragment length polymorphism were employed to detect genotypes of CAT-262C/T and CAT-21A/T polymorphism of CAT gene. Results The genotypic frequencies of CAT-262C/T and CAT-21A/T in nonintervention group,intervention group and control group were in line with Hardy-Weinberg equilibrium law (P＞ 0.05 ).The genotypes of CC and CT were detected while no TT were detected for CAT-262C/T polymorphism; the genotypes of AA, AT and TT were detected for CAT-21A/T. The genotype frequencies of CAT-262 CC, CT in control group, intervention group and non-intervention group were (89.33%(134/150), 10.67%(16/150); 88.67%(133/150), 11.33% (17/150),93.33% (140/150),6.67% (10/150), respectively. The gene frequency of C in control group, intervention group and non-intervention group were (94.67% (284/300), 94.33% (283/300),96.67%(290/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 5.33%(16/300), 5.67%(17/300), 3.33%(10/300), respectively. The genotype frequencies of CAT-21 AA,AT and TT in control group, intervention group and non-intervention group were 48.67%(73/150),46.00%(69/150),5.33%(8/150) ,52.67%(79/150) ,38.00%(57/150) ,9.33% (14/150) ,51.33%(77/150) ,38.00%(57/150), 10.67%(16/150), respectively. The gene frequency of A in control group, intervention group and non-intervention group were 71.67%(215/300),71.67%(215/300),70.33%(211/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 28.33% (85/300),28.33% (85/300),29.67% (89/300),respectively. CAT-262C/T and CAT-21A/T genotype and allele frequencies in the control group, the intervention group and non-intervention group showed no significant differences in the distribution(x2= 0.331,0.336, all P ＞0.05 ). Conclusion CAT-262C/T and CAT-21A/T polymorphism is not associated with coal-burning borne fluorosis.

ObjectiveTo observe the distribution of vitamin D receptor(VDR) gene polymorphisms in coal-burning borne fluorosis in Guizhou province and investigate the relationship between VDR gene polymorphisms and the susceptibility to coal-burning borne fluorosis.MethodsOne hundred and fifty villagers from non-improving cooking stove villages were selected as a non-intervention group in Bijie area,Guizhou province where coal-burning borne fluorosis was prevailing; 150 villagers were chosen from cooking stove improved villages as a intervention group; 150 villagers were selected from non-endemic area Changshun county as a control group.DNA was extracted from peripheral blood samples of these people.Genotype of VDR gene Bsm Ⅰ and Fok Ⅰ loci were detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).ResultsDistribution of Bsm Ⅰ polymorphism site of VDR gene of control group [AA:19.3％ (29/150),AG:39.3％ (59/150),GG:41.3％(62/150)],was compared with that[AA:4.7％(7/150),AG:14.0％(21/150),GG:81.3％(122/150)] of the non-intervention group and that[AA:7.3％(11/150),AG:23.3％(35/150),GG:69.3％(104/150)] of intervention group,and the difference was statistically significant(X2 =56.6,P ＜ 0.05).The frequency of VDR-Fok Ⅰ loci in non-intervention group [TT:29.3％(44/150),TC:55.3％(83/150),CC:15.3％(23/150)] and intervention group [TT:32.7％(49/150),TC:55.3％(83/150),CC:12.0％(18/150)] was compared with that [TT:45.3％(68/150),TC:48.7％(73/150),CC:6.0％(9/150)] of control group,and the difference was statistically significant(X2 =11.9,P ＜ 0.05).Univariate analysis showed that individuals carrying the GG genotype had increased risk of suffering fluorosis than individuals carrying the AA and AG genotypes(OR values were 6.2,3.2,all P ＜ 0.05),while carrying the TC and CC genotype had increased risk of suffering fluorosis than individuals carrying the TT genotype (OR values were 1.3,2.8,1.3,2.1,all P ＜ 0.05).ConclusionVDR gene polymorphisms may be one of the predisposing factors of coal-burning borne fluorosis.

Objective To carry on a survey on blood routine examination of coal-burning endemic fluorosis population in Bijie City,Guizhou Province in order to study their health status and problems.Methods Blood routine examination was performed in the residents in coal-fired pollution endemic fluorosis-endemic area, including the residents of the Changchun Village of Changcun Town(intervention group)whose stoves had been improved and of Shiba Village Yachi Town not improved in Bijie City,Guizhou Province.The indicators were including leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),tlle average hematocrit red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration (MCHC),red blood cell distribution width-CV(RDW-CV),platelets(PLT).Results RBC,Hb,HCT,MCHC, PLT were(4.95±1.18)×1012/L,(138.46±15.90)g/L,(50.19±11.48)%,(284.90±48.73)g/L,(334.92± 119.34)×109/L for the male in the intervened group,and they were(4.02±0.47)x 1012/L,(131.00±15.90)g/L, (40.90±7.60)%,(323.14±41.95)g/L,(280.79±100.34)× 109/L in non-intervention group,respectively. Inter-group comparison,the difference was statistically significant (U = 7.72,3.50,7.12,6.28,3.66,P ＜ 0.01). RBC, HCT,MCV,MCH,MCHC,RDW-CV,PLT were respectively(4.75±1.20)×1012/L,(46.91±11.20)%,(99.30± 6.88)fl,(28.10±8.66)pg,(275.61±54.49)g/L,(16.95±1.63)%,(351.23±150.37)×109/L for the female in the intervened group,and were (3.85±0.65)×1012/L,(38.80±6.60)%,(100.80±7.00)fl,(33.10±5.40)pg, (327.14±44.52 ) g/L,(16.60±1.58) %,(279.40±98.07)×109/L in the group un-intervened. Inter-group comparison found that there was a significant difference(U = 8.92,10.72,2.04,6.61,9.82,2.06,5.39,P ＜ 0.001 or 0.05) and the abnormal rate of RBC and Hb in non-intervention group[ 32.62% (92/282),16.67%(47/282)] was higher than that in the intervention group[9.73%(29/298) ,6.71%(20/298),x2 = 45.992,14.054,P ＜ 0.01 ) ]. Conclusion Experiment group has better results of blood routine test compared to non-intervention group,especially of anemia.

Tubulin is the major protein of microtubule. alpha- and beta- tubulins form heterodimers, while gamma-tubulin regulates microtubule organization. The present study aimed to observe the dynamic changes of gamma-tubulin in preimplantation development of parthenogenetic mouse embryos. Immunofluorescence and laser confocal microscopy were used to detect the location of gamma-tubulin in preimplantation parthenogenetic embryos activated by SrCl2. The oocytes were collected at 13-14 h after hCG injection, and then activated with 10 mmol/L SrCl2 in Ca(2+)-free CZB medium with 5 mmol/L cytochalasin B (CB), fixed at 1 h intervals until 6 h after activation. The results showed that spindle was paralleled with the cell membrane all the time, when the meiosis of MII mouse oocytes resumed. The rotation of spindle was inhibited, but karyokinesis was not influenced. At 0 h after activation, i.e. at metaphase, gamma-tubulin was distributed mainly on the two poles of spindle. At 1 h after activation, i.e. at anaphase, following the separation of chromosomes, gamma-tubulin was transformed from dense to disperse. At 2 h after activation, gamma-tubulin was localized between the segregated sister chromatids at telophase. However, at 3-6 h after activation, gamma-tubulin concentrated around the two female pronuclei during their formation and juxtaposition. Moreover, another group of MII oocytes were activated for 6 h and cultured in droplets of KSOM medium under mineral oil in 5% CO2 in air at 37 degrees C to permit parthenogenetic development. The embryos were collected and fixed at 3 h, 14 h, 16 h, and 18 h of culture. At 3 h after culture, i.e. at mitotic interphase, it was shown that amorphous gamma-tubulin distributed around the nuclei of early parthenogenetic embryos. At 24 h after culture, i.e. at prometaphase, gamma-tubulin migrated along the spindle microtubule to the two poles. Our results showed that gamma-tubulin had similar location patterns at metaphase, anaphase and telophase in meiosis and mitosis. It was concluded that gamma-tubulin assembly in parthenogenetically activated oocytes facilitated the formation of negative pole cap and the stabilization of microtubule, thus promoting the spindle formation at meiosis and mitosis. The relocation of gamma-tubulin at anaphase and telophase might be induced by the event of segregation of homologous chromosome being pulled away by the spindle. gamma-tubulin might contribute to the migration and juxtaposition of the two female pronuclei as well.
Animals ; Embryo, Mammalian ; Embryonic Development ; Female ; Meiosis ; Mice ; Mitosis ; Oocytes ; cytology ; Parthenogenesis ; Spindle Apparatus ; physiology ; Tubulin ; physiology