Adenocarcinoma ; pathology ; Aged ; Ciliary Body ; pathology ; Epithelium ; pathology ; Female ; Humans ; Uveal Neoplasms ; pathology

OBJECTIVETo discuss the changes in Wnt pathway inhibiting factors in esophageal precancerosis lesions induced by methyl benzyl nitrosamine (MBNA) and the effect of Gexia Zhuyu decoction.

METHODWistar rats were subcutaneously injected with MBNA (3.5 mg x kg(-1) for twice per week to establish the model. Since the 1st day after the model establishment, they were orally administered with Gexia Zhuyu decoction (16, 8 mg x kg(-1)). At the 10th week, esophageal tissues were collected to observe the pathological changes of esophageal mucosa, detect SFRP1, sFRP4, Axin1, Axin2 and GSK-3β mRNA levels.by fluorescent quantitation PCR analysis and β-catenin protein level by Western blotting.

RESULTBeing induced by MBNA, rats in the model group showed slight atypical hyperplasia in the histopathological examination. Compared with the normal group, Gexia Zhuyu decoction dose high and low groups showed no significant pathomorphological and histological changes. The model group showed lower gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and higher β-catenin protein expression level (P < 0.01) than the normal control group. The Gexia Zhuyu decoction low dose group showed higher gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and lower β-catenin protein expression level (P < 0.01) than the normal control group.

CONCLUSIONUp-regulated β-catenin protein level and down-regulated Wnt pathway could enhance Wnt pathway activity of MBNA-induced esophageal precancerous lesions. Gexia Zhuyu decoction could down-regulate the β-catenin protein level and up-regulate the transcription level of Wnt pathway inhibiting factors, but could not block MBNA-induced esophageal precancerosis lesions.

Animals ; Axin Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Esophageal Diseases ; drug therapy ; genetics ; metabolism ; pathology ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Male ; Necrosis ; Nitrosamines ; adverse effects ; Proteins ; genetics ; metabolism ; Rats ; Rats, Wistar ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects

The aim of this study was to observe and compare the endogenous circadian rhythm and photoresponse of Clock gene transcription in the suprachiasmatic nucleus (SCN) and pineal gland (PG) of rats. With free access to food and water in special darkrooms, Sprague-Dawley rats were housed under the light regime of constant darkness (DD) for 8 weeks (n=36) or 12 hour-light: 12 hour-dark cycle (LD) for 4 weeks (n=36), respectively. Then, their SCN and PG were dissected out every 4 h in a circadian day, 6 rats at each time (n=6). All animal treatments and sampling during the dark phases were conducted under red dim light (<0.1 lux). The total RNA was extracted from each sample and the semi-quantitative RT-PCR was used to determine the temporal mRNA changes of Clock gene in the SCN and PG at different circadian times (CT) or zeitgeber times (ZT). The grayness ratio of Clock/H3.3 bands was served as the relative estimation of Clock gene expression. The experimental data were analyzed by the Cosine method and the Clock Lab software to fit original results measured at 6 time points and to simulate a circadian rhythmic curve which was then examined for statistical difference by the amplitude F test. The main results are as follows: (1) The mRNA levels of Clock gene in the SCN under DD regime displayed the circadian oscillation (P<0.05). The endogenous rhythmic profiles of Clock gene transcription in the PG were similar to those in the SCN (P>0.05) throughout the day with the peak at the subjective night (CT15 in the SCN or CT18 in the PG) and the trough during the subjective day (CT3 in the SCN or CT6 in the PG). (2) Clock gene transcription in the SCN under LD cycle also showed the circadian oscillation (P<0.05), and the rhythmic profile was anti-phasic to that under DD condition (P<0.05). The amplitude and the mRNA level at the peak of Clock gene transcription in the SCN under LD were significantly increased compared with that under DD (P<0.05), while the value of corresponding rhythmic parameters in the PG under LD were remarkably decreased (P<0.05). (3) Under LD cycle, the circadian profiles of Clock gene transcription induced by light in the PG were quite different from those in the SCN (P<0.05). Their Clock transcription rhythms were anti-phasic, i.e., showing peaks at the light phase ZT10 in the SCN or at the dark time ZT17 in the PG and troughs during the dark time ZT22 in the SCN or during the light phase ZT5 in the PG. The findings of the present study indicate a synchronous endogenous nature of the Clock gene circadian transcriptions in the SCN and PG, and different roles of light regime in modulating the circadian transcriptions of Clock gene in these two central nuclei.
Animals ; CLOCK Proteins ; genetics ; Circadian Rhythm ; physiology ; Male ; Photoreceptor Cells, Vertebrate ; physiology ; Pineal Gland ; physiology ; Rats ; Rats, Sprague-Dawley ; Suprachiasmatic Nucleus ; physiology ; Transcription, Genetic

OBJECTIVETo identify the anatomical variability of the left spermatic vein (LSV) and determine its effect on the induction of experimental left varicocele (ELV) in adolescent rats.

METHODSWe equally randomized 30 adolescent male SD rats to groups A (LSV collaterals fully ligated and the left renal vein constricted), B (only the left renal vein constricted), and C (sham operation), observed the courses of the LSVs and measured their diameters. At 30 days after operation, we analyzed the changes in the left kidneys and the diameters of the LSVs.

RESULTSIrregular collaterals were observed in 90% of the LSVs and no abnormal changes were found in the left kidneys after surgery. The postoperative LSV diameter was remarkably increased in group A as compared with the baseline ([1.47 +/- 0.15 ] vs [0.16 +/- 0.08] mm, P < 0.01), but showed no significant difference in group B ([0.31 +/- 0.49] vs [0.15 +/- 0.07] mm, P > 0.05) and C ([0.17 +/- 0.07] vs [0.16 +/- 0.06] mm, P > 0.05), and it was significantly longer in A than in B (P < 0.01). The success rate of ELV induction was 100% in group A and 10% in group B, but no varicocele was observed in group C.

CONCLUSIONCorrect identification of the anatomical course of the LSV and ligation of its irregular collaterals are essential for the establishment of a stable and consistent ELV model.

Animals ; Disease Models, Animal ; Kidney ; pathology ; Ligation ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord ; blood supply ; Varicocele ; Veins ; abnormalities

The effects of different physical and chemical factors on hairy root growth and flavonoids production were studied in suspension culture of Saussurea medusa hairy root in 1/2 MS medium. The results showed that the following culture conditions, nitrogen concentratiaon (involved NH4+ and NO3-), 30 mmol/L; the ratio of ammonium to nitrate, 5:25; the combination of 2% sucrose and 3% glucose; 0.5 mg/L GA3; 0.5 mg/L IBA; initial pH 5.8; light cycle, 18 h/d (3500lx); temperature, 24 degrees C; shaker revolutions per minute, 100 r/min, were favourable to hairy root growth and flavonoids production. Under the above culture conditions, up to 12.8 g/L (DW) of hairy root and 1922 mg/L of flavonoids were obtained after 21 days of culture. The content of total flavonoids in hairy root was 15%, which was about 25 times as that in the wild plantlet.
Culture Media ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; metabolism ; Saussurea ; growth & development ; metabolism ; Temperature ; Tissue Culture Techniques

Objective To observes the change of early effective biomarkers of endothelial injury with lowarsenic exposure in drinking water. MethodsNinety rurad residents, who had lived in Yanhe village, Xuyi county and Jiangsu province for at least 10 years, were recruited by simple random sampling in this study. The level of arsenic in their household shallow well were divided into three groups, which were ＜ 10 (32 people), 10 - 50(28 people) and ＞ 50 μg/L(30 people). Blood samples from individuals were collected. Malondialdehyde(MDA) in human plasma, which is considered as the most important marker for monitoring lipid peroxidation, was determined as conjugate with tetrabutylammonium hydrogen sulphate(TBA). The level of anti-superoxide anion radical(O-·2),C-reactive protein(CRP) and NO in human plasma was measured with colorimetry, turbidimetry and nitric acid reductase, respectively. The number of circulating endothelial progenitor cells(CEPCs) in peripheral blood was analyzed by CD133+/KDR+ antibodies and flow cytometry. Results Ninety cases underwent questionnaires. Between the groups, the difference of the levels of MDA (61.1, 65.5, 67.5 μmol/kg), O-·2 (4774.6, 5143.3, 4736.0 U/kg) ,CRP[(5.92 ± 2.44), (5.11 ± 2.40), (5.55 ± 2.96)mg/L], and NO[(659.8 ± 387.5), (667.4 ± 486.6), (762.1 ±763.2)μmol/kg], was not statistically significant (F =0.00, 0.46, 0.80, 0.47, all P ＞ 0.05). The difference of the number of CEPCs in different groups of arsenic in drinking water was statistically significant(0.96 x 10-5, 0.77 x 10-5,1.59 x 10-5, F=5.08, P＜ 0.05), where ＜ 10, 10 - 50 μg/L groups were significantly lower than ＞ 50 μg/L group (q =4.58, 6.65, all P ＜ 0.05). ConclusionsThe number of CEPCs in peripheral blood changes significantly with lower-arsenic exposure, whereas there are no obvious changes with the markers of oxidized damage and inflammation. This is the first human demonstration showing that lower-arsenic exposure may cause endothelial injury.

OBJECTIVETo study the protective effects of salidroside on injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cell.

METHODApoptosis and intracellular free calcium concentration ([Ca2+]i) were measured by flow cytometry, morphological changes and neuronal necrosis were observed with fluorescence microscope, and the lactic dehydrogenate (LDH) release was measured by LDH kits.

RESULTHypoxia/hypoglycemia cultures for 2 hours induced neuronal apoptosis and necrosis. They were 18.59% (P < 0.01) and 4.94% (P < 0.01) respectively. There were morphological changes including chromatin condensation, nuclear fragmentation and formed apoptotic bodies after exposed to hypoxia/hypoglycemia for 2, 4, 6, 12 hours. After 2 hours of hypoxia/hypoglycemia, neuronal [Ca2+]i and the release of LDH were significantly increased. They were 8.46 nmol/L (P < 0.01) and 16.59% (P < 0.01) respectively. The effects were enhanced with the extending time of hypoxia/hypoglycemia. Salidroside might have significantly decreased the percentage of neuronal apoptosis and necrosis, reduced neuronal [Ca2+]i and the release of LDH. The effects of salidroside were strengthened with the increasing of Salidroside dosage.

CONCLUSIONSalidroside has effect of anti-neuronal apoptosis. This effect might be related to its function of decreasing intracellular free calcium concentration.

Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Hypoglycemia ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Phenols ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhodiola ; chemistry

Spermatogenesis is a complex regulatory process depending on a variety of hormones (such as FSH, LH, T, and 17beta estradiol), cytokines, and genes. Research on gene regulation in spermatogenesis has become a hot spot and revealed some spermato-genesis-related genes, such as AYZ, DAZ, YRRM, NOSTRIN, and so on. Reports are rarely seen on the role of CR16 in male reproduction, and its action mechanism in spermatogenesis is not yet clear. This article updates the role of CR16 in spermatogenesis in the male reproductive system from the perspective of Sertoli cells forming a blood-testis barrier.
Animals ; Blood-Testis Barrier ; Humans ; Male ; Microfilament Proteins ; Sertoli Cells ; Spermatogenesis ; Testis ; cytology

The present study investigated the influence of the methyl jasmonate and salicylic acid elicitors on the formation of phenylethanoid glycosides (PeG) in the suspension cultures of Cistanche deserticola. The results showed that methyl jasmonate and salicylic acid enhanced greatly the accumulation of PeG and echinacoside (Echin), but their optimum elicitation dosage and addition time were different. The yields of PeG and Echin were significantly increased in the presence of 5 micromol/L methyl jasmonate on day 14 (up to 2.59-fold and 3.82-fold, respectively), whereas treated with 50 micromol/L salicylic acid on day 28, the maximum content of them were, respectively, 2.71 and 3.16-fold higher than the untreated cell cultures.
Acetates ; pharmacology ; Cell Culture Techniques ; Cistanche ; drug effects ; metabolism ; Culture Media ; Cyclopentanes ; pharmacology ; Glycosides ; biosynthesis ; Oxylipins ; pharmacology ; Phenylethyl Alcohol ; metabolism ; Salicylic Acid ; pharmacology

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P