HK-2 cells were cultured with various concentrations of glucose and insulin for 12,24,48,72 h.Transforming growth factor-?_1(TGF-?_1) protein in supematant was measured by ELISA,while TGF-?_1 mRNA expression was assessed by RT-PCR.Data showed that high concentration of glucose and insulin up-regulated the expression of TGF-?_1 in HK-2 cells through different pathways.

1.0?108 co-pies/L)and the lower viral load group(HBV DNA

ObjectiveTo investigate the long term effect of different antihypertensive agents on renal hemodynamics with ultrasound Doppler.Methods52 essential hypertensive patients were divided into three groups according to the antihypertensive agent they took: angiotensin-converting-enzyme inhibitor (ACEI), calcium channel blocker (CCB) and β-adrenoceptor blocker (βB). The blood pressures of right upper arms were measured with mercury column blood pressure gauge using Korotkoff method. Renal hemodynamics were examined with ultrasound Doppler before and after treatment with agents. ResultsSystolic blood pressure (SBP) and diastolic blood pressure (DBP) of all patients were decreased significantly. An significant correlation was found between the deceases of blood pressure and the betterment of renal homodynamic parameters in ACEI and CCB groups, but not in βB group. ConclusionTo a degree, the betterment of renal hemodynamics is correlation with the reduction of systemic blood pressure and mechanism of action of antihypertensive agents.

OBJECTIVETo analyze the mutation of the KAL1 gene in male patients with idiopathic hypogonadotropic hypogonadism (IHH).

METHODSWe analyzed the exon mutation of the KAL1 gene in 30 IHH patients using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with the PCR product direct sequencing technique.

RESULTSThree cases of the KAL1 gene mutation were found among the total number of patients, including 1 case of nonsense mutation (c. 1270C > T,p. R424X), and 2 cases of frameshift mutation, (c. 279_280delAG,p. G94fs) and (c. 1886_1887delTT,p. L629fs).

CONCLUSIONOf the 3 cases of the KAL1 gene mutation we detected, 2 are new and 1 already reported in the literature. The results of our study have provided valuable information on the molecular genetics of the IHH syndrome.

Adolescent ; Adult ; Base Sequence ; Child ; DNA Mutational Analysis ; Exons ; Extracellular Matrix Proteins ; genetics ; Humans ; Hypogonadism ; genetics ; Kallmann Syndrome ; genetics ; Male ; Mutation ; Nerve Tissue Proteins ; genetics ; Polymorphism, Single-Stranded Conformational ; Young Adult

OBJECTIVE Oxidative sress is one of the key factor responsible for occurrence and development of hepatic fibrosis, a common consequence of chronic liver injury of multiple etiology. Nuclear factor erythroid 2-related factor 2 (Nrf2) serves as a major regulator of a celular defense system against oxidative stress. Xiaochaihutang (XCHT), a compound of seven botanical extracts used for liver diseases traditionally in East Asia. However, few studies have investigated its anti-hepatic fibrosis effects and pathophysiological mechanism of action. The present study was designed to confirm the anti-hepatic fibrosis effects and explore its potential mechanism of action by investigating the intervention of Nrf2 pathway. METHODS Liver fibrosis was induced by repeated injection of Carbon tetrachloride (CCl4) over a period of 9 weeks. Starting from the 6th week, the animals in treatment groups were given the appropriate dose of XCHT granules and silybin. Biochemical parameters, histological changes of the liver and alpha-smooth muscle actin (α-SMA) were determined. The expressions of Nrf2, Keap1, Nqo1, HO-1, Gclc and Gclm were assessed by RT-PCR and Western blot. RESULTS CCl4 caused a significant fibrosis damage in the rat liver and the liver functions and fibrosis degree were significantly improved by XCHT (5 g·kg- 1 and 10 g·kg- 1). XCHT (5 g·kg- 1 and 10 g·kg- 1) treatment significantly decreased the number of cells labeled with α-SMA antibodies. Moreover, XCHT (5 g·kg-1 and 10 g·kg-1) significantly increase Nqo1, HO-1, Gclc and Gclm expressions in the liver. CONCLUSION These studies establish XCHT is a potentially useful therapeutic agent for treatment of hepatic fibrosis and it might be via regulation of Nrf2 pathway in rats against oxidative stress, making further efforts to inhibiting the activated HSCs. Activation or up-regulation of Nrf2 pathway may be an alternative treatment strategy for liver fibrosis.

BACKGROUND:The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injuryin vivo in a mouse model of sepsis.METHODS:Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.RESULTS:Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3-4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.CONCLUSIONS:Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.

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 determine the mRNA and protein expressions of RCAS1 in human astrocytic tumors, and to explore the relation between their expression and the genesis and development of tumor. METHODS: The RCAS1 mRNA expression in human astrocytic tumors was evaluated by RT-PCR, and the RCAS1 protein expression was studied by immunohistochemical staining. RESULTS: The quantities of RCAS1 mRNA expression between diffusive astrocytoma(Grade II) and anaplastic astrocytoma(Grade III), anaplastic astrocytoma and glioblastoma(Grade IV) were significantly different(P<0.05), while the expression scores of RCAS1 protein were different only between the anaplastic astrocytoma and glioblastoma(P<0.01). RCAS1 protein expression was positively correlated with the tumor grade (r=0.573,P<0.001). The RCAS1 protein was not detected in normal brain tissues by immunohistochemical staining. CONCLUSION The RCAS1 expression is related to the histological grade of astrocytic tumor. In astrocytic tumors, the RCAS1 expression is regulated transcriptionally and posttranscriptionally.
Adolescent ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; metabolism ; Astrocytoma ; genetics ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma (HCC). In this study, we investigated the anti-proliferative activities and anti-angiogenesis effects of saikosaponins (SS)-b on hepatocellular carcinoma (HCC) and its regulation on VEGF/ERK/HIF-1α signal pathway. METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro. Pathological change of tumor tissue was observed by HE staining, the microvascular changes were detected by immunohistochemical method. The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane (CAM) model. The effects of SS- b on proliferation, migration and invasion were investigated by MTT assay, scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell (HUVEC) and HepG2 cells in vitro. Vascular endothelial growth factor (VEGF), matrix metalloproteinase-2/9(MMP-2/9), hypoxia-inducible factor-1α (HIF-1α) expression and the phosphorylation of extracellular regulated kinase(ERK) were analyzed using RT-PCR and Western-blot. RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo. The inhibitory rate of tumor was 49.1%, 50.7%, 66.1% in SS-b 5, 10 and 20 mg·kg-1 group respectively. HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice. Moreover, SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF. SS-b had an obvious inhibitory effect on cell proliferation, migration and invasion of HUVEC cells and HepG-2 cells. These effects were associated with down-regulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1α signaling in H22 mice and Hep-G2 cells. CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibit?ing tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.

AIM:The effect of acupuncture on mitophagy-related protein expression in skeletal muscle of rats after heavy-load exercise was investigated to explore the role of acupuncture in the repairment of exercise-induced skeletal muscle damage. METHODS:Male adult Sprague-Dawley rats (n=128) were randomly divided into 4 groups:control (C,n=8) group, exercise (E, n=40) group, acupuncture (A, n=40) group, and exercise and acupuncture (EA, n=40) group. The rats in E group and EA group performed an eccentric exercise,and the rats in A group and EA group immediately after exercise received acupuncture treatment. The rats in the latter 3 groups were further divided into 0 h,12 h,24 h,48 h and 72 h sub-groups(n=8),and soleus muscle was collected at each time point. The transmission electron microscopy was used to observe the ultrastructural changes of the mitochondria in skeletal muscle. The content of citrate synthase (CS) was measured by ELISA. The protein expression of skeletal muscle PTEN-induced putative kinase 1 (PINK1),parkin and microtubule-associated protein 1 light chain 3 (LC3) was determined by Western blot. RESULTS:After the heavy-load exercise,the mitochondria swelled and accumulated under cell membrane. The number of mitophago-somes was increased,and the content of CS was significantly decreased(P<0.05). The expression of PINK1,parkin and LC3 was significantly elevated (P<0.05). However,the acupuncture intervention after exercise promoted the recovery of mitochondrial ultrastructure, attenuated mitophagolysosome formation, maintained CS content and down-regulated the ex-pression of PINK1,parkin and LC3 (P<0.05). CONCLUSION:Heavy-load exercise causes the damages of mitochon-drial structure and function in the skeletal muscle and activates PINK1/parkin pathway to induce excessive occurrence of mitophagy. Acupuncture intervention after exercise is able to alleviate the damage of mitochondria in the skeletal muscle through decreasing the expression of mitochondrial outer membrane protein PINK1,reducing the recruitment of downstream cytoplasmic protein parkin,thereby affecting the combination of LC3 and mitochondria to inhibit the overactivation of mito-phagy.