Objective To analyze genetic and clinical features of 14 children with ?-thalassemia intermedia in Guangxi area.Methods ?-thalassemia genes,?-thalassemia genes,single nucleotide polymorphism(SNP) at position-158 of()~G?-globin gene,AT repeats polymorphisms of DNase I-hypersensitive site 2 of the ?-globin gene cluster locus control region(?-LCR-HS2) were detected by PCR techniques.Clinical data were analyzed.Results Genotype:1.Seven cases were homozygous or compound heterozygous for nt-28(A→G).Among them,2 cases′ genotypes were nt-28/nt-28,1 case was ?~E/ nt-28,2 cases were ?~0/nt-28,1 case(?~0/nt-28) co-inherited()~G?158(T) and 1 case(?~0/nt-28) co-inherited simultaneously()~G?-158(T) and——SEA ?-thalassemia-1 genes.2.Three cases with ?~0/?~0 presented()~G?-158(T),and other 3 cases co-inherited——SEA ?-thalassemia-1 genes.3.One patient with ?~0/?~0 co-inherited()~G?-158(T) and——SEA ?-thalassemia-1 genes.4.Six cases carrying()~G?-158(T) had(AT)_9 N_(12)(AT)_(10) sequences in ?-LCR-HS2.Phenotype:The values of Hb,MCV,HbF of 14 patients were(75.9?9.7) g/L,(68.9?5.9) fL,66.9%?16.3%,respectively.Except for 2 cases with genotypes of nt-28/nt-28 and 1 case with ?~E/nt-28 who had never been transfused,the others had more severe symptoms and required irregularly transfusion.Conclusions In the 14 children with ?-thalassemia intermedia from Guangxi area,nt-28(A→G),()~G?-158(T) and——SEA ?-thalassemia-1 gene are main alleviating gene factors.Incidence of(AT)_9 N_(12)(AT)_(10) sequence in ?-LCR-HS2 in these patients is high.Patients who are homozygous for nt-28 or compound heterozygous for ?~E have milder phenotypes.

OBJECTIVETo investigate the estrogenic activity of icariin and genistein with estrogen-dependent human breast cancer (MCF-7) cells.

METHODMCF-7 cells were incubated with media containing 5% charcoal dextran-treated FBS in phenol red-free media for 48 h. CCK-8 kit was used to study the impact of defferent concentration of icariin and genistein on MCF-7 proliferation in vitro. Optimal concentration icariin and genistein were added into medium and total RNA was isolated after 12, 24, 36, 48 h. The gene expression of ERalpha, ERbeta, PS2, and PR were investigated by Real-time RT-PCR Total protein was also isolated and secretion of ERalpha, ERbeta, PS2, and PR were examined by Western blot.

RESULT10 micromol x L(-1) icariin and genistein could promote the proliferation of MCF-7 evidently. However, the ability of genistein to promote the proliferation was better than icariin. With the concentration of 10 micromol x L(-1), genistein group had a stronger expression of ERa, PS2 and PR mRNA levels than icariin while ERbetaexpression had no significant difference in two group. The same effects were detected by western blotting.

CONCLUSIONBoth genistein and icariin have a strong estrogen-like effect, but the estrogenic activity of genistein is stronger than icariin. It showed that the activity of icariin is stron-ger than genistein to promote ROB maturation. So it must be that icariin promotes the maturation of osteoblasts in vitro by a estogen-independent mechanism.

Cell Proliferation ; drug effects ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Estrogens ; pharmacology ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; Genistein ; pharmacology ; Humans ; MCF-7 Cells ; Osteoblasts ; cytology ; drug effects ; metabolism ; Presenilin-2 ; metabolism

In the reductive branch, glycerol is first dehydrated to 3-hydroxypropionaldehyde that is then reduced to 1,3-PD under the consumption of reducing power (NADH). If over-expression of the gene dhaB encoding glycerol dehydrase is achieved,the reducing power will be scarce and 3-hydroxypropionaldehyde will be accumulated,which is disadvantage to produce 1,3-propanediol.The structure gene yqhD from E.coli encoding 1,3-propanediol oxidoreductase isoenzyme[under the consumption of reducing power (NADPH)]and the gene dhaB encoding glycerol dehydrase from Klebsiella pneumoniae was amplified using PCR method.The two gene were transferred into expression vector pEtac to construct a novel recombinant Klebsiella pneumoniae (pEtac-dhaB-tac-yqhD).Over-expression of yqhD and dhaB in Klebsiella pneumoniae was achieved with pEtac-dhaB-tac-yqhD.The fermentation result on aerobic conversion showed the increase of 20% of 1,3-propanediol yield by Klebsiella pneumoniae(pEtac-dhaB-tac-yqhD) was obtained compared with Klebsiella pneumoniae.The main by-products,acetic acid and butanediol decreased estrogen receptors 30% obviously.

Objective This objective of the study was to analyze the factors related to being recaptured and condom use among low-fee female sex workers (FSWs) to provide reference in developing human immunodeficiency virus (HIV) intervention strategy. Methods Physical examination certificates were designed by Zhongshan County Center for Disease Control and Prevention to record HIV and syphilis test results for low-fee FSWs from 2013 to 2015. Low-fee FSWs were asked to show physical examination certificates in the next intervention and test. Multivariate Logistic regression was used to analyze factors associated with being captured with physical examination certificates. Generalized linear mixed model was used to analyze factors associated with condom use with clients. Results A total of 220 low-fee FSWs were recruited by using physical examination certificates and received 389 interviews from 2013 to 2015. The proportions of HIV positive and syphilis positive were 4.2% (9/213) and 30.0% (64/213) respectively among those who had HIV and syphilis test. Results of multivariate Logistic regression analysis showed that low-fee FSWs who had been FSWs for more than 4 years (OR=2.95, 95% CI：1.35-6.45), and worked in the local county in the past 30 days (OR=11.74, 95% CI: 5.26-26.20), were more likely to be captured with physical examination certificates. Results of generalized linear mixed model showed that those who were captured at least once (OR=3.33, 95% CI: 1.34-8.27), had junior middle school education and above (OR=22.79, 95% CI: 3.75-138.57), had high HIV knowledge (OR=3.57, 95% CI: 1.52-8.38), and charged more than 30 yuan for vaginal sex (OR=30.68, 95% CI: 12.57-74.90), were more likely to use condom consistently. Conclusions Physical examination certificates could be used for low-fee FSWs surveillance and intervention and tracking their HIV and syphilis status. The intervention strategy should take these into consideration.

OBJECTIVETo investigate the sensitivity of gastric cancer stem cells (GCSCs) to 5-fluorouracil (5-FU), and to explore the cytological mechanism of drug-resistance of gastric cancer.

METHODSImmunohistochemical staining was employed to examine expression of stem cell marker CD44 and drug-resistance protein thymidylate synthase (TS) in 71 cases with gastric cancer. Based on morphology of cell colonies derived from AGS gastric cell line, colonies consisting of GCSCs were isolated, and expression of CD44 and TS, as well as self-renewal capacity of GCSCs were detected. Sensitivity of GCSCs to 5-FU was examined through CCK-8 assay.

RESULTSThe positive rates of CD44 and TS were 59.2% (42/71) and 56.3% (40/71) in gastric cancer, and expression of CD44 was associated with that of TS (χ(2)=12.76, P<0.01; Kappa=0.41). Serial sections indicated that CD44+ cancer cells simultaneously expressed TS. AGS developed morphologically diverse colonies, and the GCSCs colonies exhibited a tight and regular shape, which were called holoclone. Holoclones expressed CD44 and TS strongly, possessing capacity of robust self-renewal and forming a lot of second passage clones after incubation. Subclones expressed CD44 and TS weakly, forming less second passage clones. Paraclones almost did not express CD44 and TS, forming no second passage clone after incubation. Affected by 5-Fu, three holoclones showed less growth inhibition compared with another colony type and wild-type AGS cells. Furthermore, IC50 of 5-FU for three holoclones was (113.43±5.81), (272.68±25.75) and (118.14±17.75) μmol/L respectively, which were significantly higher than that of one subcolony type [(16.97±1.01) μmol/L] and AGS cells [(27.52±0.59) μmol/L] (all P<0.05).

CONCLUSIONSGCSCs possess lower sensitivity to 5-FU, and may play a critical role in drug-resistance of gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Humans ; Male ; Middle Aged ; Neoplastic Stem Cells ; drug effects ; pathology ; Stomach Neoplasms ; pathology ; Tumor Cells, Cultured ; Young Adult

In order to find the cardiotonic constituents of lateral roots of Aconitum carmichaelii Debx., the investigation was carried out. Silica gel column chromatography, Sephadex LH-20, medium-pressure MCI and reverse phase ODS column chromatography were used to separate the 90% EtOH extract of the lateral roots of Aconitum carmichaelii Debx. The structures of the isolated compounds have been identified by chemical properties and spectroscopic analyses. Ten compounds were isolated and their structures were elucidated as benzoic acid-5-hydroxy-2-benzoyl-amino methyl ester (1), honokiol (2), pinoresinol (3), salicylic acid (4), p-hydroxy-cinnamic acid (5), songorine (6), karakoline (7), mesaconitine (8), hypaconitine (9) and 14-benzoylhypaconitine (10), separetely. Compound 1 is a new compound and its structure has been established by NMR, HR-ESI-MS, UV, IR and X-Ray. Compound 2-5 are isolated from the lateral roots of Aconitum carmichaelii Debx. for the first time.
Aconitum ; chemistry ; Cardiotonic Agents ; chemistry ; isolation & purification ; Plant Roots ; chemistry

Objective To compare the effects of moxibustion at different temperatures on blood lipid and TRPV1 in dorsal root ganglion with hyperlipidemia rats; To verify the correlation between efficacy of adjusting fat of moxibustion with activating of TRPV1. Methods The rat model of hyperlipidemia was made by high fat diet. 60 SD mice were randomly divided into four groups: control group, model control group, the 38 ℃ moxibustion group and the 45 ℃ moxibustion group, with 15 mice in each group. Acupoints Shenque and Zusanli were chosen under moxibustion for 10 minutes each time, once another day, for 4 weeks, in the 38 ℃ moxibustion group and the 45 ℃moxibustion group. Blood was taken after intervention, and blood TC, TG, LDL-C and HDL-C of the mice were detected by oxidase method; the dorsal root ganglion was taken to detect the expression of TRPV1 mRNA by real-time fluorescence quantitative PCR. Results Compared with model group, blood lipid indexes in moxibustion groups had different changes, with statistical significance compared with 45 ℃ moxibustion group (P<0.05, P<0.01), and there was statistical significance between 38 ℃ moxibustion group and 45 ℃ moxibustion group (P<0.01); there was statistical significance in TRPV1 mRNA of dorsal root ganglion among 45 ℃ moxibustion group and other three groups (P<0.01). Conclusion The correlation between efficacy of adjusting fat of moxibustion with activating of TRPV1 has been confirmed.

To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established.
Naphthaleneacetic Acids ; pharmacology ; Phenylurea Compounds ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Shoots ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Thiadiazoles ; pharmacology ; Tissue Culture Techniques ; Tulipa ; drug effects ; growth & development

To test the safety of using cardiac death donation ( DCD ) corneas for penetrating keratoplasty surgery graft.METHODS:ln chronological order, suing DCD corneas penetrating keratoplasty, corneal endothelial cell density and best corrected visual acuity ( BCVA) were tested 3~4mo after surgery.RESULTS:A total of 14 cases of DCD while 26 corneas were included in this study. Donors age ranged 0. 5 ~61 years, averagely 38. 3 ± 15. 6 years. Causes of death included that 9 cases of traumatic brain injury, 2 cases myocardial infarction, 2 cases brain stem hemorrhage, 1 case of respiratory and circulatory failure. All 26 patients underwent penetrating keratoplasty, no rejection occurred and all grafts were transparent 3 ~ 4mo after surgery. Three to four months after surgery, corneal endothelial cell density ranged 794 ~ 4 347/mm2 , averaged 2 305 ± 827/mm2 , within which was only one case was lower than 1000/mm2 (3. 8%), while 9 cases ranged from 1000 ~ 2000/mm2 (34. 6%), 16 cases were higher than 2000/mm2 (61. 5%). The age of all the 26 receipts were from 20~80 years, mean 40. 7±17. 1 years. BCVA before surgery was light perception positive to 0. 08, with an average 0. 027±0. 024. Three to four months after surgery, BCVA were 0. 2~0. 8, with an average 0. 52± 0. 182 in contrast (t=3. 96, P<0. 001).CONCLUSlON:DCD donated corneas could be used for penetrating keratoplasty graft with high security.

Objective To compare the effects of invasive blood pressure monitoring of the shallow temporal artery and radial artery in infants. Methods 40 infants with serious illness were randomly selected to monitor their blood pressure by puncturing the shallow temporal artery and radial artery on the same side. The success rate of puncture at a time, average puncture time, press time after extubation and complications during cannulation were compared. Results The success rate of puncture at a time in the shallow temporal artery was higher than that in the radial artery ( x2 = 12.17 ,P ＜0. 01 ). The average puncture time for the shallow temporal artery and the radial artery were ( 7.05 ± 1.50) min and ( 11.18 ± 2.51 ) min, respectively, and the differences were statistically significant( t = 11.69, P ＜ 0. 01 ). The press time of the shallow temporal artery after extubation [(9.45 ± 1.72) min]was longer than that of the radial artery[( 8.00 ± 1.63 ) min]( t = 10. 84, P ＜ 0. 01 ),the occurrence of needle displacement and dislocation during cannulation were significantly higher in the shallow temporal artery ( x2 = 4.501, P ＜ 0. 05 ) and the occurrence of hemorrhage and hematoma at the puncture site was not significantly different (x2 = 0. 353,0. 215;P ＞ 0. 05 ). Conclusions It is safe and feasible to choose the shallow temporal artery for invasive blood pressure monitoring in infants with serious illness, and it could improve the success rate of puncture, save puncture time, not only gain time for rescue, but also reduce nurses' workload.