OBJECTIVETo explore the mechanism of Astragalus polysaccharides (APS) for improving the cardiac function of Sjogren's syndrome (SS) model rats based on Keapl-Nrf2/ARE signaling pathway.

METHODSTotally 48 male Wistar rats were randomly divided into four groups by random digit table, i.e., the blank control group,the model control group,the APS group, and the hydroxychloroquine group, 12 in each group. Except those in the blank control group, 0. 1 mL mixed antigen protein of sufficiently emulsified Freund's complete adjuvant and submandibular gland protein was injected from two feet plantar to induce SS model. The intervention was started from 19th day after inflammation induction. Equal volume of normal saline was given to rats in the blank control group (1 mL/100 g), APS was administered to those in the APS group (1 mg/100 g), and hydroxychloroquine (0.03 125 g/kg) was administered to those in the hydroxychloroquine group. All rats were intervened once per day for 30 consecutive days. Changes of rats' body mass and drinking water quantity, submandibular gland index, spleen index, histological changes of glands were observed. Changes of the heart function were monitored using invasive hemodynamics. Serum reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity (TAC), tumor necrosis factor alpha (TNF-alpha), and interleukin-35 (IL-35)were detected using ELISA method. The pathological changes were observed using HE staining. The protein expression of ROS, reactive nitrogen species (RNS), glutathione (GSH), and thioredoxin (TRX) were observed by immunohistochemical staining. The mRNA expression of Keap1, Nrf2, and ARE was detected using real time fluorescent quantitative PCR. The protein expression levels of gamma-glutamic acid and a half long glycine synthetase (gamma-GCS) and heme oxygenase 1 (HO-1) in the myocardial tissue were determined by Western blot method. Results Compared with the blank control group, the quantity of drinking water, submandibular gland index, spleen index, heart rate (HR), cardiac index (HI), left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVEDP), MDA, ROS, TNF-alpha, ROS protein expression, RNS protein expression, Keap 1 mRNA expression, Maf mRNA expression, Nfr2 mRNA expression, and HO-1 protein expression, and gamma-GCS protein expression significantly increased (P <0.01); body mass, +/-dp/dtmax, SOD, TAC, IL-35, GSH, and TRX significantly decreased (P <0.01) in the model group. Compared with the model group, the quantity of drinking water, submandibular gland index, spleen index, LVEDP, MDA, ROS, TNF-alpha, ROS protein expression, RNS protein expression, Keap1 mRNA expression, Maf mRNA expression, Nfr2 mRNA expression, and HO-1 protein expression, and gamma-GCS protein expression significantly decreased (P<0.05); body mass, +/-dp/dtmax, SOD, TAC, IL-35, GSH protein expression, and TRX protein expression significantly increased (P < 0.05, P <0.01) in the AR group and the hydroxychloroquine group. In the hydroxychloroquine group HR increased (P <0.05). In the AR group HR and LVSP decreased (P <0. 05, P <0. 01). Compared with the hydroxychloroquine group, HR, LVEDP, - dp/dtmax, y-GCS protein expression significantly decreased (P <0. 05, P <0. 01); SOD, TAC, GSH, TRX, HO-1 protein expression increased (P <0.01 )in the AR group. HI was positively correlated with ROS (P <0. 05). LVSP and LVEDP were positively correlated with Keap1 -Nrf2/ARE signaling pathways (P <0. 01) , and negatively correlated with TAC (P <0. 05, P <0. 01 ). +/-dp/dtmax was negatively correlated with Keap1-Nrf2/ARE signaling pathways(P <0.05), and positively correlated with TNF alpha (P <0. 05).

CONCLUSIONSDeclined heart function exists in SS rats. The mechamechanism of APS for improving the heart function might be closely correlated with activating Keap1-Nrf2/ARE signaling pathway.

Animals ; Astragalus Plant ; Blotting, Western ; Carboxylic Ester Hydrolases ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hydroxychloroquine ; Male ; Malondialdehyde ; metabolism ; Myocardium ; enzymology ; NF-E2-Related Factor 2 ; metabolism ; Plant Extracts ; therapeutic use ; Polysaccharides ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Sjogren's Syndrome ; Submandibular Gland ; Superoxide Dismutase ; metabolism ; Thioredoxins ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective　To explore the generative mechanism of bacterial wave growth in view of the open living system of microorganism. Methods　With the single strain of bacterium as the subject, the nonlinear method of the dynamic systems and the analyzing method of the numerical simulation were used to study the effect of the changing environment on the bacterial growth. Results　A mathematical model of the relationship between the bacterial wave growth and environment was established. Conclusion　This model is verified to be scientific and its numerical simulation results are obtained, providing a helpful quantitative analysis for the research on the laws of bacterial wave growth.

Objective To establish an animal model by placing one end of PICC in the hepatic portal vein of a beagle dog and leaving the other end out of its body.Methods Six Beagle dogs were given respiration anesthesia through orotracheal intubation.An incision was made through the right rectus abdominalis to locate the superior mesenteric vein (SMA) and the main hepatic portal vein.The left branch of SMA was separated and cut to put PICC into the main hepatic portal vein before being ligated and fixed.The other end of PICC was elicited through the right abdominal wall and passed beneath the skin to the back neck and fastened in case of movement.Results The anesthetic effect was good and all the operations were successful.The mean operation time was about an hour and the mean blood loss was about 15 ml.The incision healed 5-7 d after operation.Conclusion The establishment of the model can improve the effects of liver-targeting drugs,which can cut down the dosage,lower the cost of treatment and experiment and reduce the adverse effect of medicines.Through PICC,we can directly draw blood from the hepatic portal vein to measure the blood concentration before the first pass elimination.Then according to the concentration,we can calculate the absorption rate in the gastrointestinal tract,which can facilitate related experimental studies.

OBJECTIVETo explore whether there is phenotypic modulation of mesangial cells in streptozotocin (STZ) induced diabetic rats and study the effect of Tujian Mixture (TJM) on it.

METHODSSD rats were divided into the normal control group (NC group, n = 8), the unilateral nephrectomized control group (QC group, n = 8), the STZ induced diabetes mellitus with unilateral nephrectomy model group (DM group, n = 8), the Valsartan treated group (VT group, n = 8) and the TJM treated group (ZY group, n = 9), rats in the latter two groups were modeled as in the DM group and treated with Valsartan (20 mg/kg.d) and TJM (20 g/kg.d) respectively for 12 weeks. The expression of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor-beta 1 (TGF-beta 1) in rats glomeruli were observed by immunohistochemistry assay, and the ratio of alpha-SMA and TGF-beta 1 positive area/total glomerule tuft area (SMA/GT and TGF/GT) were analyzed using computer-assisted image analysis software.

RESULTSIn the NC and the QC groups, only trace of alpha-SMA positive staining was found. But there was prominant alpha-SMA positive staining in glomeruli of the DM group, with SMA/GT and TGF/GT increased significantly (P < 0.01), and marked increase of 24 hrs proteinuria excretion (P < 0.01). As compared with the DM group, the three indexes were all significantly lower in the VT and ZY groups (P < 0.01), and the lowering of proteinuria was more significant in the ZY group than that in the VT group (P < 0.01).

CONCLUSIONThe expression of alpha-SMA in glomeruli in STZ induced diabetic rats with unilateral nephrectomy is pronounced, indicating that phenotypic modulation of mesangial cells involvement in the pathogenesis of diabetic nephropathy. TJM and Valsartan can reduce 24 hrs proteinuria excretion, inhibit the phenotypic modulation of mesangial cells and the expression of TGF-beta 1 in glomeruli of diabetic rats, and the effect of TJM is more potent than that of Valsartan in lowering urinary protein excretion.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetic Nephropathies ; etiology ; metabolism ; pathology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Glomerular Mesangium ; metabolism ; pathology ; Image Processing, Computer-Assisted ; Male ; Nephrectomy ; Phenotype ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo examine the changes of expressions of orexin A, orexin receptor-1 (OX1R), prepro-orexin (Prepro-OX) mRNA, OX1R mRNA and ob-R of hypothalamus in rats with chronic renal failure (CRF).

METHODSSixty-two male Wister rats weighing 200-250 g were divided into three groups, including group 1 (normal, n = 5), group 2 (sham-operated, n = 25) and group 3 (CRF, n = 32). Hypothalamus orexin A was assayed by radioimmunoassay. Serum leptin was assayed by enzyme linked immunosorbent assay. The expression of Prepro-OX mRNA and OX1R mRNA of hypothalamus were measured by reverse transcription polymerase chain reaction, and expression of orexin A, OX1R and ob-R by immunohistochemistry. Automatic biochemical analyzer was used to measure the serum creatinine.

RESULTSHypothalamus orexin A levels were negatively correlated (r = -0.63, P < 0.001) with serum leptin levels in the rats. The expression of hypothalamus Prepro-OX mRNA in CRF rats was significantly lower than that of sham-operation at week 12 (P < 0.01). Hypothalamus Prepro-OX mRNA levels were negatively correlated (r = -0.81, P < 0.001) with the levels of serum leptin and serum creatinine (r = -0.68, P < 0.05) in the rats at week 12. The expression of hypothalamus OX1R mRNA in CRF rats was lower than that of sham-operation at week 12 (P > 0.05). Specific immunoreactivity for orexin A was present in perikeryon of the hypothalamus neuron. Specific OX1R-like immunoreactivity was observed in some nerve fibres. Specific immunoreactivity for ob-R was present in membranes of the hypothalamus neuron. Hypothalamus neurons of orexin A-like specific immunoreactivity in CRF rats were significantly fewer than those in shamoperated rats at week 8. Hypothalamus neurons of OX1R-like specific immunoreactivity in CRF rats were similar to those in sham-operated rat at week 8. Hypothalamus neurons of ob-R-like specific immunoreactivity in CRF rats were significantly more than those in sham-operated rats at week 8.

CONCLUSIONSThe lower hypothalamus orexin A levels may be induced by high serum leptin level in CRF rats. The lower expression of hypothalamus Prepro-OX mRNA in CRF rats may be one of the main causes inducing lower hypothalamus orexin A. The expression of OX1R in hypothalamus neurons is somewhat reduced and the expression of ob-R in hypothalamus neurons is somewhat raised in CRF rats. These remain to be studied further.

Animals ; Carrier Proteins ; genetics ; metabolism ; Hypothalamus ; metabolism ; Intracellular Signaling Peptides and Proteins ; Kidney Failure, Chronic ; metabolism ; Leptin ; genetics ; metabolism ; Male ; Neuropeptides ; genetics ; metabolism ; Neurotransmitter Agents ; genetics ; metabolism ; Orexin Receptors ; Orexins ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Cell Surface ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; Receptors, Leptin ; Receptors, Neuropeptide ; genetics ; metabolism

OBJECTIVETo observe the effect of bone morphogenetic protein-7 (BMP-7) on the transdifferentiation of cultured human tubular epithelial cell (HKC) induced by TGF-beta1 and to elucidate its possible mechanism.

METHODSThe cultured HKC cells were divided into 5 groups: serum-free group (negative control); single TGF-beta1 treated group (positive control); single BMP-7 treated group; combined TGF-beta1 and BMP-7 treated group; and BMP-7 pre-treated group. Expression of keratin of HKC cells was assessed by indirect enzyme immunohistochemistry (IEI), expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin by immunohistological method, percentage of alpha-SMA positive HKC cells by flow cytometry, and mRNA expression of alpha-SMA, TGF-beta1, and TGF-beta type II receptor by reverse transcription PCR.

RESULTSThe expression of alpha-SMA and the percentage of alpha-SMA positive HKC cells markedly increased after having been treated by TGF-beta1 while the expression of E-cadherin and keratin decreased. In the group pre-treated with BMP-7 (50 ng/ml) and then added with TGF-beta1 (8 ng/ml), expression of alpha-SMA was significantly lower than in the positive control group, while expression of E-cadherin and keratin significantly higher than in the positive control group. Measurement of the percentage of alpha-SMA positive HKC found significant deference between the combined TGF-beta1 and BMP-7 treated group and the positive control group (9.7% vs 19.8%; 5.8% vs 19.8%; P < 0.05). Significant difference existed between the BMP-7 (50 ng/ml) pre-treated group and the positive control group (8.7% vs 19.8%, P < 0.05). mRNA expression of alpha-SMA was measured by RT-PCR and the results showed that it significantly decreased in the group treated or pre-treated with BMP-7 (50 ng/ml) (15% and 12% of the results in the positive control group, respectively). The mRNA expression levels of both TGF-beta1 and its type II receptor significantly decreased (28% and 19%; 47% and 36%, compared with the positive control group, respectively).

CONCLUSIONTransdifferentiation of cultured renal epithelial cell induced by TGF-beta1 can be inhibittd by certain levels of BMP-7, cultured together with TGF-beta1 or pretreated. BMP-7 can prevent and inhibit the mRNA expression of TGF-beta1 and its type II receptor, which may be an important mechanism by which BMP-7 inhibit the transdifferentiation of renal tubular epithelial cell.

Actins ; biosynthesis ; genetics ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules ; cytology ; metabolism ; Polymerase Chain Reaction ; RNA, Messenger ; biosynthesis ; genetics ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1

OBJECTIVETo examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1).

METHODSThe transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot.

RESULTSThe immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners.

CONCLUSIONSIncreased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

Cell Differentiation ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules, Proximal ; cytology ; RNA, Messenger ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo examine the relationship between effect of vascular endothelial growth factor (VEGF) on epithelial-myofibroblast transition (EMT) of HK-2 cells and changes in expressions of bone morphogenetic protein-7 (BMP-7) and inhibitor of DNA binding/differentiation (Id) 2, Id3.

METHODSThe cultured HK-2 cells were co-treated with transforming growth factor-beta1 (TGF-beta1) (5 ng/ml) and VEGF165 (0.1, 1, 10, 100 ng/ml), or with TGF-beta1 (5 ng/ml) and VEGF receptor-1 neutralized antibody (10 microg/ ml), and were also co-treated with TGF-beta1 (5 ng/ml) and VEGF165 (100 ng/ml) with or without activin receptor-like kinase 6 (Alk6)/Fc Chimera (2 microg/ml, to neutralize endogenous BMP-7) for 48 hours. mRNA and protein expressions of alpha-smooth muscle actin (alpha-SMA), E-cadherin, BMP-7, Id2 and Id3 of HK-2 cells were assessed with double-stain immunocytochemistry, real-time PCR and Western blot respectively.

RESULTSCompared with normal controls, alpha-SMA expression significantly increased, while E-cadherin, BMP-7, Id2, and Id3 mRNA and protein expressions markedly decreased in HK-2 cells treated with TGF-beta1 (5 ng/ml) (P < 0.05). VEGF165 interrupted TGF-beta1 induced alpha-SMA expression in a dose-dependent manner and upregulated BMP-7, Id2 mRNA and protein expressions of the cells (P < 0.05). alpha-SMA expression increased, while E-cadherin, BMP-7, and Id2 expressions decreased further in HK-2 cells co-treated with TGF-beta1 and VEGFR1 antibody compared with normal controls (P < 0.05). When endogenous BMP-7 was neutralized with Alk6/Fc Chimera in the cells co-treated with TGF-beta1 and VEGF165, alpha-SMA expression upregulated (P < 0.05), while Id2 was not changed.

CONCLUSIONSVEGF165 may partially inhibit TGF-beta1-induced EMT of HK-2 cells in vitro. This effect is related to the upregulated expressions of BMP-7 and Id2. Id2 may be upregulated directly by VEGF165, but not related to BMP-7.

Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; Cell Line ; Epithelial Cells ; cytology ; metabolism ; Gene Expression Regulation ; Humans ; Inhibitor of Differentiation Protein 2 ; genetics ; metabolism ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVEPituitary prolactinoma with severe erectile dysfunction (ED) as the initial symptom is often misdiagnosed. This article explores the diagnosis and treatment of severe ED caused by pituitary prolactinoma.

METHODSWe retrospectively analyzed the diagnosis and treatment of 4 cases of pituitary prolactinoma with severe ED (IIEF-5 score 5 - 7) as the initial clinical symptom confirmed by MRI.

RESULTSThe 4 cases of pituitary prolactinoma-induced severe ED, with serum prolactin 10 times above the maximum normal level, were misdiagnosed for 2 years. All failed to respond to the PDE5 inhibitor therapy, and then 3 of them underwent transnasal hypophysectomy. Twenty-four months of follow-up found the level of prolactin restored to normal in 1 case (IIEF-5 = 19), and reduced to 600 and 768 IU/L respectively (IIEF-5 = 15) in the other 2. Then administration of the PDE5 inhibitor was followed, which produced satisfactory efficacy. One case was treated with oral bromocriptine, which restored the prolactin level to normal at 12 months (IIEF-5 > 21).

CONCLUSIONProlactin detection and brain MRI can help to confirm pituitary prolactinoma with severe ED at the onset. As for its treatment, in case of an extremely high level of prolactin, simple administration of the PDE5 inhibitor is ineffective. When the prolactin level is reduced after surgery or medication, the symptom of ED can be improved and, in case of no obvious relief, administration of the PDE5 inhibitor can be followed, which may achieve satisfactory results.

Adult ; Erectile Dysfunction ; diagnosis ; etiology ; Humans ; Male ; Middle Aged ; Phosphodiesterase 5 Inhibitors ; therapeutic use ; Pituitary Neoplasms ; complications ; diagnosis ; drug therapy ; Prolactinoma ; complications ; diagnosis ; drug therapy ; Retrospective Studies

OBJECTIVETo evaluate surgical outcomes and the feasibility of robotic thyroidectomy and central neck dissection (CND).

METHODSThe clinical data of 40 patients of papillary thyroid microcarcinoma underwent total thyroidectomy (or lobectomy and isthmusectomy) and CND using the Da Vinci system through axillo-bilateral-breast approach in Jinan Military General Hospital of People's Liberation Army from February to December 2014 were analyzed retrospectively (robotic group). Other forty patients of papillary thyroid microcarcinoma underwent total thyroidectomy (or lobectomy and isthmusectomy) and CND by open approach were selected as the control (open group). Cosmetic satisfaction was assessed after a month postoperation by the numerical score system. t-test and χ(2) test were used to compare the clinical characters, total operative time, intraoperative estimated blood loss, postoperative hospital stay, number of lymph nodes removed, visual analogue scale for pain, postoperative complications, and cosmetic effect between the 2 groups.

RESULTSAll 80 patients were diagnosed of papillary thyroid microcarcinoma. The total thyroidectomy (or lobectomy/isthmusectomy) with CND of 40 patients were successfully performed by da Vinci Si surgical system. The numbers of total thyroidectomy of robotic group and the open group were 36 and 37, respectively. The numbers of metastatic lymph nodes of robotic group and open group were 14 and 15, respectively. The operation time of the robotic group was (130±12) minutes, which was longer than that of open group (98±11) minutes (t=12.432, P<0.05). The study showed statistical significant difference between the two groups regarding the visual analog scale pain assessment (1.9±0.9 vs.3.9±1.1, t=8.900, P<0.05). There were no statistical significant difference of intraoperative estimated blood loss, postoperative hospital stay, number of lymph nodes removed, and the complication rate between the 2 groups.Postoperative cosmetic result was more satisfying on the robotic group (9.1±0.5) than open group (4.8±1.5) (t=17.200, P<0.05).

CONCLUSIONSThe robotic total thyroidectomy (or lobectomy and isthmusectomy) and CND has similar surgery safety and feasibility as open procedures. The robotic thyroidectomy is a good alternative surgical modality for patients with papillary thyroid microcarcinoma who wish to avoid neck scars.

Axilla ; Breast ; Carcinoma, Papillary ; surgery ; Humans ; Length of Stay ; Lymph Nodes ; Neck Dissection ; Operative Time ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Robotic Surgical Procedures ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; methods