OBJECTIVETo explore the toxic effect of fluoride on the human thyroid cells (Nthy-ori 3-1) and its mechanism.

METHODSNthy-ori 3-1 cells were exposed to 0.0, 0.1, 1.0, 3.0 mmol/L of sodium fluoride (NaF) in vitro. After 24 hours incubation, 3 (4,5-Dimethylthiazol-z-yl)-3, 5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay were used to measure cell viability and the LDH leakage rate. Reactive oxygen species (ROS) level, constituent ratio of the cell cycle, and apoptosis rate were measured by flow cytometry.

RESULTSComparing to viability of control group (set as 100.00%), the cell viability of the 1.0, 3.0 mmol/L fluoride-treated groups (76.64 +/- 9.13)%, (64.04 +/- 6.32)% were significantly decreased (all P values <0.01). LDH leakage rate and ROS level of the 3.0 mmol/L fluoride-treated group ((48.66 +/-7.15)%, (29993.50 +/- 1786. 86) FI) were significantly increased (all P values <0.01) compared to control group ((35.24 +/- 3.02)%, (13021.33 +/- 1067.55) FI). The G0/G1 phase cells of the 1.0 mmol/L fluoride-treated group ((40.76 +/- 5.65)%) were lower than control group (60.09 +/- 1.76)% (P < 0.01), yet the percentage of cells in S phase ((54.05 +/- 4.59)%) were higher than the control group (32.59 +/- 2.43) % (P < 0.01). Comparing to control group ((9.64 +/- 3.44)%), the percentage of apoptosis cells increased in the 3.0 mmol/L fluoride-treated group ((20.09 +/- 3.22)%) (P < 0.01).

CONCLUSIONTo Nthy-ori 3-1 cells, fluoride under experimental concentrations decreases cell viability, improve the LDH leakage rate, and ROS level. It blocks the cells in S phase and induce cell apoptosis.

Apoptosis ; drug effects ; Cell Cycle ; Cell Division ; Cell Line ; Fluorides ; toxicity ; Humans ; Reactive Oxygen Species ; analysis ; Thyroid Gland ; cytology ; drug effects

The output and agronomic characters of 3-year-old Panax notoginseng cultured under stereo structure (upper, middle and down layers) were investigated, and the annual change of N, P and K of its planting soil were also studied. Results showed that, compared with field cultured Panax notoginseng, growth vigour and output of stereo-cultivation were significantly lower. But the total production of the 3 layers was 1.6 times of field. The growth vigor and production of P. notoginseng was in the order of upper layer > middle layer > down layer. The content of ginsenoside in rhizome, root tuber and hair root of P. notoginseng was in the order of upper layer > field > middle layer > down layer. Organic matter content and pH of stereo-cultivation soil decreased with the prolonging of planting time, which with the same trend of yield. Organic matter content of stereo-cultivation soil was significantly higher than field, but the pH was significantly lower. Contents of total and available N, P and K in stereo-cultivation soil and field decreased with the prolonging of planting time. The content of N and P were in the order of upper layer > middle layer > yield > down layer, the content of K was in the order of upper layer > middle layer > down layer > yield. Compared with field, the proportion of N and P in the organ of underground (rhizome, root tuber and hair root) of upper layer were increased, while decreased in middle and down layers. Proportion of K in underground decreased significantly of the 3 layers. In conclusion, the agronomic characters and production of stereo-cultivation were significantly lower than that of yield. But the total production of the 3 layers were significantly higher than field of unit area. And the aim of improving land utilization efficiency was achieved. Nutritions in the soil of stereo-cultivation were enough to support the development of P. notoginseng, which was not the cause of weak growth and low production. The absorbing ability of P. notoginseng to N, P and K nutrients was decreased by stereo-cultivation mode. So, improve the growth vigour of P. notoginseng from the perspective of adjusting the stereo-cultivation mode so as to improve the nutrient absorption capacity is the future direction.
Food ; Hydrogen-Ion Concentration ; Nitrogen ; analysis ; Panax notoginseng ; growth & development ; Phosphorus ; analysis ; Potassium ; analysis ; Soil ; chemistry

AIMTo explore the protective effects of ischemic preconditioning on the kidney injury following with ischemia/reperfusion (I/R) of limbs.

METHODSThe models of I/R injury of limbs were constructed in rabbits. The blood from right external jugular vein, renal artery and renal vein represent the peripheral blood, into and out-flowing kidney blood (IKB, OKB) respectively. Superoxide dismutase (SOD), malondialdehyde (MDA), blood uria nitrogen (BUN) in peripheral blood and SOD, MDA, nitric oxide (NO) in IKB and OKB were measured, as well as SOD, MDA, induced nitric oxide synthase (iNOS) in kidney were detected in different groups. The effects of ischemic preconditioning (IPC) on the kidney injury were observed.

RESULTSCompared with control group, the activity of SOD in peripheral blood, IKB, OKB and kidney decreased, and the content of MDA increased after 4 h ischemia followed by 4 h reperfusion. The content of BUN in peripheral blood, NO in IKB, OKB and iNOS in kidney increased remarkably as well. SOD increased and MDA, NO, BUN, iNOS decreased significantly by ischemic preconditioning (IPC) before ischemia/reperfusion. The correlation analysis indicated that MDA was negatively correlated with SOD and positively correlated with NO, BUN.

CONCLUSIONOxygen free radicals metabolic confusion of kidney occurred in the course of I/R of limbs, IPC could strengthen the resistance of peroxidation in kidney and had protective effects on the kidney injury following with ischemia/reperfusion (I/ R) of limbs

Animals ; Blood Urea Nitrogen ; Extremities ; blood supply ; Female ; Ischemic Preconditioning ; methods ; Kidney ; metabolism ; physiopathology ; Lipid Peroxidation ; physiology ; Male ; Rabbits ; Renal Insufficiency ; etiology ; prevention & control ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

BACKGROUND:WWOX,a tumor suppressor gene,can affect the growth of ovarian cancer stem cells;however,there is no report on whether its mechanism of action is related to Hedgehog signaling pathway.OBJECTIVE:To investigate the effect and mechanism of overexpression of WWOX on the apoptosis of ovarian cancer stem cells.METHODS:pcDNA3.1-WWOX (pcDNA3.1-WWOX group) and pcDNA4.0-WWOX (pcDNA4.0-WWOX group) were transferred into ovarian cancer stem cells,respectively;and meanwhile,pcDNA3.1 (pcDNA3.1 group) and pcDNA4.0 (pcDNA4.0 group) were transferred into the cells.A non-transfection group (only with Lipofectamine2000) was set up.After cultured 48 hours,the levels of WWOX in the pcDNA3.1-WWOX group and pcDNA4.0-WWOX group were detected using western blot assay,and the cell proliferation and apoptosis were detected using MTT assay and flow cytometry,respectively.Western blot assay was also used to detect the levels of Hedgehog signaling pathway associated proteins,SHH,PTCH1,Gli-1,SMO and apoptosis-related protein Cleaved Caspase-3 in the cells.Cyclopamine,Hedgehog signaling pathway inhibitor,was used in ovarian cancer stem cells without transfection (cyclopamine group) and after the transfection of WWOX overexpression vector (WWOX+cyclopamine group) followed by 48 hours of culture,and then MTT,flow cytometry and western blot detections were performed.RESULTS AND CONCLUSION:(1) The expression level of WWOX in the pcDNA4.0-WWOX group was significantly higher than that in the pcDNA3.1-WWOX group (t=27.84,P=0.00).The ovarian cancer stem cells which were transfected with pcDNA4.0-WWOX were used to overexpress WWOX in the late experiment.(2) Overexpression of WWOX could inhibit the proliferation of ovarian cancer stem cells and promote the apoptosis of ovarian cancer stem cells.(3) Overexpression of WWOX could inhibit the expression of Gii-1,PTCH1,SMO and SHH in ovarian cancer stem cells,and promote the expression of Cleaved Caspase-3.(4) Cyclopamine could inhibit the expression of SHH,PTCH 1,Gli-1,SMO,and promote the expression of Cleaved Caspase-3.Cyclopamine had obvious inhibitory effect on Hedgehog signaling pathway.(5) Cyclopamine could enhance the apoptosis induced by overexpression of WWOX in ovarian cancer stem cells,and enhance the inhibition of proliferation of ovarian cancer stem cells induced by overexpression of WWOX.To conclude,WWOX effects on proliferation and apoptosis of ovarian cancer stem cells may be related to the inhibition of Hedgehog signaling pathway.

BACKGROUND: Opportunistic infection of Candida albicans (C. albicans) has become a serious problem in immunocompromised patients. The study aimed to explore the mechanism of enterogenous infection of C. albicans in immunocompromised rats under severe acute pancreatitis (SAP). METHODS: Sprague Dawley (SD) rats (n=100) were randomly assigned into 5 groups as the following: blank group, cyclophosphamide+ceftriaxone+SAP group, cyclophosphamide+ceftriaxone group, cyclophosphamide+SAP group, and cyclophosphamide group. The rats were sacrificed at 5 and 10 days, and their jejunum, colon, mesenteric lymph nodes, pancreas, intestinal content, and blood were quickly collected to detect C. albicans. A region of the 25S rRNA gene was chosen and amplified by polymerase chain reaction (PCR) to differentiate C. albicans genotypes. The amplified products were further sequenced and compared to judge their homology. RESULTS: Compared with the Cyclophosphamide group, the combination of immunosuppressants and broad-spectrum antibiotics significantly increased the colonization of C. albicans in intestine in 5 and 10 days. Pure SAP stress did not increase the opportunistic infection of C. albicans. The PCR products of C. albicans isolates all belonged to the genotype A family, and sequence alignment showed that the amplified fragments were homologous. CONCLUSION: The damage of immune system and broad-spectrum antimicrobial agents are important risk factors for opportunistic fungal infection. Intestinal tract is an important source for genotype-A C. albicans to translocate and invade into bloodstream.

A laccase gene (lacD) from the basidiomycete Trametes sp. 420 was heterologously expressed in Pichia pastoris in two ways, resulting in two recombinant enzymes of rLacDx with native N-terminus and rLacDe with eight additional amino acid residues at N-terminus. The yields of rLacDx and rLacDe in shaken-flask cultures after an 18-day growth were 1.21 x 10(5) u/L and 7.38 x 10(4) u/L, respectively, as determined with 2,2'-azinobis(3-ethylbenzothia-zoline- 6-sulfonic acid) (ABTS) as substrate. The yield of rLacDx was further increased to 2.39 x 10(5) u/L under high-density fermentation while the production process was decreased to 7.5 days. In addition, rLacDx and rLacDe exhibited similar enzymatic characters in oxidizing substrate guaiacol, and were stable at 50 degrees C and at a pH range from 3 to 10. However, the specific activity of rLacDx (1761 u/mg) for ABTS was higher than that of rLacDe (1122 u/mg), and the apparent Km value of rLacDx (427 microM) was less than that of rLacDe (604 microM).
Cloning, Molecular ; Fermentation ; Isoenzymes ; biosynthesis ; genetics ; Laccase ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Trametes ; enzymology ; genetics

OBJECTIVETo investigate the role of D-galactose, especially in the structural and functional changes of the immune system in aging.

METHODSSerum levels of advanced glycation end-products (AGE) were determined by ELISA method. Ultra-structures of thymus and spleen were detected by transmission electron microscopy. MTT method was used to determine the lymphocyte proliferation. IL-2 activity was determined by bioassay. Northern blot was used to detect the IL-2 mRNA levels.

RESULTSSerum AGE levels of D-galactose- (P < 0.01) and AGE-treated (P < 0.05) mice (n = 8) were increased significantly. The ultra-structures of thymus and spleen in D-galactose- and AGE-treated mice showed regressive changes similar to those in the aged control group. The lymphocyte mitogenesis and IL-2 activity of spleen were also decreased significantly (P < 0.01, n = 8). The change of IL-2 activity shown by Northern blot resulted from the change of mRNA expression. The AGE plus aminoguanidine group, however, showed no significant change in these parameters in comparison with the young control group (P < 0.01 or P < 0.05, n = 8).

CONCLUSIOND-galactose and AGE lead to a mimic regression change of aging in the immune system in vivo.

Aging ; drug effects ; immunology ; Animals ; Cell Proliferation ; drug effects ; Galactose ; pharmacology ; Glycation End Products, Advanced ; blood ; Interleukin-2 ; metabolism ; Lymphocytes ; drug effects ; immunology ; Mice ; Microscopy, Electron, Transmission ; RNA, Messenger ; metabolism ; Spleen ; drug effects ; immunology ; ultrastructure ; Thymus Gland ; drug effects ; immunology ; ultrastructure

OBJECTIVETo investigate association between the natural-resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms and susceptibility to pulmonary tuberculosis (TB) in Chinese Han population.

METHODSHospital-based case-control study design was adopted. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) technique were used to type three NRAMP1 polymorphisms (INT4, D543N and 3'UTR). Information on related factors of tuberculosis was collected using a pre-tested standard questionnaire. Univariate and multivariate unconditional logistic analyses were conducted using SPSS for window software package. Totally, 110 cases of TB were selected during April 2001 to June 2002, with an average age of (27.7 +/- 12.7) years. Also, 180 cases of healthy control were selected, aged (27.3 +/- 9.2) years in average. Locus of NRAMP1 polymorphism was analysed with univariate method.

RESULTSUnivariate analysis demonstrated that the D543N G/A and 3'UTR TGTG+/del genotype occurred more frequently in the cases than in the controls, with crude odds ratios (OR) (95% CI) of 2.22 (1.03 - 4.78) and 1.93 (1.14 - 3.26), respectively. No significant association was observed between TB and INT4 polymorphisms. In multivariate analysis, associations of TB and D543N G/A and 3'UTR TGTG+/del genotypes remained, adjusted for exposure history and bacille Camette-Guérin immunization. Adjusted OR (95% CI) was 3.04 (1.12 - 8.27) and 2.36 (1.20 - 4.64), respectively. Still, no significant association between INT4 polymorphisms and TB was found.

CONCLUSIONPolymorphisms of D543N and 3'UTR locus in NRAMP1 gene might affect their susceptibility to TB in Chinese Han population.

Adolescent ; Adult ; Case-Control Studies ; Cation Transport Proteins ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Logistic Models ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Tuberculosis, Pulmonary ; genetics

OBJECTIVETo observe the therapeutic effect of tranquilizing and allaying excitement needling method on insomnia of heart and spleen deficiency and the effect of brain blood flow.

METHODSSixty cases were randomly divided into a tranquilizing and allaying excitement needling method group (observation group) and an eight confluence points selected group (control group), 30 cases in each group. The observation group was treated by acupuncture at Sishencong (EX-HN 1), Shenmen (HT 7), and Sanyinjiao (SP 6) with tranquilizing and allaying excitement needling method. The control group was treated by acupuncture at Shenmai (BL 62) and Zhaohai (KI 6). Their therapeutic effects and changes of brain blood flow were observed.

RESULTSThe total effective rate was 93.3% (28/30) in the observation group which was better than 83.3% (25/30) in the control group (P < 0.05). After treatment, the peak velocity of systolic (Vp) and diastolic blood flow velocity (Vd) of middle cerebral artery, basilar artery and vertebral artery were increased in the both groups (P < 0.01, P < 0.05), with more obvious increase in the observation group (P < 0.01, P < 0.05), except the Vd of left vertebral artery.

CONCLUSIONBoth the tranquilizing and allaying excitement needling method and the eight confluence points selected needling method can improve the clinical signs and symptoms. Meanwhile, the therapeutic effect of acupuncture is related with improvement of brain blood flow. However, the tranquilizing and allaying excitement needling method has better therapeutic effect on insomnia of heart and spleen deficiency.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Brain ; blood supply ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Spleen ; physiopathology ; Young Adult

OBJECTIVETo evaluate the effectiveness of the video-assisted minimally invasive radiofrequency ablation combined irbesartan use for the treatment of the persistent atrial fibrillation (AF).

METHODSFrom January 2006 to December 2009, 83 patients with persistent AF having a video-assisted minimally invasive radiofrequency ablation. There were 58 males, 25 females with a mean age of (57 ± 11) years. Mean duration of preoperative AF was (61 ± 65) months. Follow-up for the whole patients ranged from 1.0 to 3.6 years [mean (2.2 ± 0.8) years]. Patients were randomly divided into irbesartan group (n = 42) and without irbesartan group (n = 41) postoperatively.

RESULTSNo patient died postoperatively. During follow-up, there was 1 patient died of unknown reason. At the end of the procedure, 38 patients (45.7%) were sinus rhythm, 4 patients (4.9%) were pacing rhythm, 5 patients (6.0%) were atrial flutter or atrial tachycardia, and 36 patients (43.4%) were AF. Before discharge, 53 patients (63.9%) were sinus rhythm, 24 patients (28.9%) were AF. At late follow-up, 65 patients (80.2%) were sinus rhythm; 14 patients (17.3%) were AF or atrial flutter. After follow-up, the Kaplan-Meier analysis showed the irbesartan group had fewer patients with AF (P = 0.020). The hazard ratio for AF recurrence in patients treated with irbesartan was 0.24 (95% CI: 0.087 to 0.637, P = 0.004).

CONCLUSIONSThe video-assisted minimally invasive radiofrequency ablation is safe and effective. The patients treated with irbesartan have a lower rate of recurrence of AF.

Adult ; Aged ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Proportional Hazards Models ; Retrospective Studies ; Thoracic Surgery, Video-Assisted ; Treatment Outcome