Aim To observe the effect of mycophenolate mofetil(MMF) on expressions of MCP-1 and CD68 in renal tubulointerstitial injury of diabetic rats and explore the mechanism of MMF′s protective role.Methods Diabetes was induced in uninephrectomized male Wistar rats by peritoneal injection of STZ (65 mg?kg-1). Rats were randomly divided into three groups:control group (NC),diabetic group (DM) and treated group (DM+MMF) with MMF(15 mg?kg-1?d-1).This study lasted for 8 weeks. 24 h urinary protein,blood glucose and the ratio of left kidney weight/body weight were determined after 8 weeks.The renal tubulointerstitial morphological change was observed,immunohistochemical method was used to analyze expressions of MCP-1 protein and CD68. Expression of MCP-1 mRNA in renal tissue was measured by quantitative Real-time PCR.Results Compared with NC group, serum glucose level,24 hour urinary protein and the ratio of left kidney weight/body weight were significantly increased(P

A comparable study were carried out by determination of trace elements on five marine-derived shell traditional Chinese medicine (TCM) (Ostreae Concha, Haliotidis Concha, Margaritifera Concha, Meretricis Concha, and Arcae Concha), which were recorded in the Chinese Pharmacopoeia (2010 version). Seven trace elements in 51 batches of this type of shell TCM were analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), combined with principal component analysis (PCA) methods. The content of element Se, which exhibited significant differences among different drugs, could be used as a key element to distinguish this type of drugs. Meanwhile, the contents of elements Co, Cu, Mo, and Ba in Haliotidis Concha, Co and As in Margaritifera Concha, Mo and As in Meretricis Concha, Mo, As, and Ba in Arcae Concha, and Zn in Meretricis Concha were relatively stable. In the PCA plot, Arcae Concha and Meretricis Concha could be efficiently distinguished from Ostreae Concha together with Haliotidis Concha, and Margaritifera Concha. The results also showed a correlation with their medicinal function. In conclusion, trace elements in marine-derived shell TCM could not be neglected for their quality control.
Animal Shells ; chemistry ; Animals ; Aquatic Organisms ; chemistry ; Bivalvia ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Trace Elements ; analysis

BACKGROUND: Previous research has demonstrated that dermal tissue has mesenchymal stem cells, which have a possibility of autologous transplantation. If the mesenchymal stem cells derived from the skin differentiate into lymphocytes under a certain condition, the immune system disease can be solved generally.OBJECTIVE: To investigate the possibility of differentiation of human skin-derived mesenchymal stem cells into lymphocytes. METHODS: Surface marker expression was detected in the 14th passage human skin-derived mesenchymal stem cells using flow cytometry. Transdifferentiation medium of human skin-derived mesenchymal stem cells consisted of human lymphocyte supernatant and fresh human skin-derived mesenchymal stem cells based on the ratio of 7:3. Inverted microscope was employed to observe morphological changes, and flow cytometry was used to detect surface marker expression in the lymphocytes at 1-8 days after induction. Self-marker expression of human skin-derived mesenchymal stem cells was then detected at 3,6, and 9 days after induction.RESULTS AND CONCLUSION: Human skin-derived mesenchymal stem cells stably expressed self-specific marker CD73, Vimentin and so on, but did not express specific markers of hematopoietic system, I.e., CD34, CD45 and so on, lowly expressed HLA-I, but did not express HLA-DR at all. At 3 days after induction, the cell volume significantly increased, cell proliferation rate was significantly lower than before induction, and a lot of cystic-like particles with strong refraction were observed in or between cells. The CD45 lymphocyte expression was not significantly changed, but CD3, CD19, CD16, CD4, and CD8 expression rates of human skin-derived mesenchymal stem cells were linearly increased at 1-4 days after induction and stabilized at 5-8 days after induction. In addition, CD37, CD34, Vimentin, and HLA-DR expressions were not changed at 3, 6, and 9 days after induction, but HLA-I expression rate was gradually increased with the prolongation time of induction. This suggested that human skin-derived mesenchymal stem cells can differentiate into lymphocyte and potentially participate in repairing immune system injury.

BACKGROUNDPodocyte has inflammatory role in the development of diabetic nephropathy (DN). Mycophenolate mofetil (MMF), an anti-inflammatory agent, can suppress macrophage infiltration and reduce renal injury in streptozotocin-induced diabetic rats. Angiotensin II receptor blocker (ARB), another renal protecting agent, can decrease podocyte loss in DN. In this study, we detected the expression levels of monocyte chemoattractant protein-1 (MCP-1) and nephrin to evaluate podocyte's role in inflammatory reaction in DN, observe and compare the effect of MMF alone and in combination with valsartan, on preventing podocyte loss in streptozotocin (STZ) induced diabetic rats.

METHODSDiabetic model was constructed in uninephrectomized male Wistar rats by single peritoneal injection of STZ (65 mg/kg). The successfully induced diabetic rats were randomly divided into four groups: diabetes without treatment group (DM), valsartan treated group (DMV), MMF treated group (DMM), and combined therapy group (DMVM). Normal rats of the same sibling were chosen as control (NC). At the end of the 8th week, serum biochemistry, 24-hour urinary protein (UP) and the ratio of kidney weight/body weight (RWK/B) were measured. The rats were sacrificed for the observation of renal histomorphology through light and electron microscope. Nephrin, desmin and MCP-1 levels were detected by semi-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1.

RESULTSCompared with group NC, serum glucose level, 24-hour UP and RWK/B in group DM were significantly higher (P < 0.01), and the nephrin mRNA level in DM group was significantly lower (P < 0.05). The nephrin mRNA expression levels in group DMV, DMM and DMVM were all higher than that of DM group (P < 0.05) and no significant differences were found among the three treatment groups (P > 0.05). Treatment with MMF, valsartan or their combination could significantly decrease the 24-hour UP and RWK/B, and suppress glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. In diabetic rats, the high expressions of desmin and MCP-1 in kidney were suppressed by valsartan, MMF or their combination.

CONCLUSIONSPodocytes are involved in the inflammatory reaction of diabetic rats. MMF could suppress MCP-1 and desmin expression, enhance nephrin expression, and attenuate proteinuria in diabetic rats. The combined therapy of valsartan and MMF did not show any superiority over monotherapies on renal protection. MMF may have renoprotective effect in early stages of diabetic nephropathy through preventing podocytes loss and anti-inflammatory activity.

Animals ; Chemokine CCL2 ; analysis ; Desmin ; analysis ; Diabetic Nephropathies ; drug therapy ; pathology ; Drug Therapy, Combination ; Immunohistochemistry ; Male ; Membrane Proteins ; analysis ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; therapeutic use ; Podocytes ; drug effects ; pathology ; Rats ; Rats, Wistar ; Tetrazoles ; administration & dosage ; therapeutic use ; Valine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Valsartan

AIMTo explore the modulation of 5-HT on GABA-activated current (I(GABA)) in the membrane of rat dorsal root ganglion (DRG) neurons and its mechanism.

METHODSRat DRG neurons were isolated mechanically and enzymatically, on which whole-cell patch clamp recording and repatch technique for intracellular dialysis were performed.

RESULTSIn the majority of neurons examined (92.0%, 69/75) GABA induced a concentration-dependent inward current. In neurons sensitive to GABA preapplication of 5-HT produced potentiation effect (82.6% , 57/69) on I(GABA). Preapplication of 5-HT at concentrations of 1 x 10(-6), 1 x 10(-5), 1 x 10(-4) and 1 x 10(-3) mol x L(-1) potentiated I(GABA) by (35 +/- 8)% (n=8), (47 +/- 11)% (n=10), (65 +/- 17)% (n=9) and (75 +/- 18)% (n=11), respectively. This effect was mimicked by alpha-methyl-5-HT (1 x 10(-6) mol x L(-1)), a specific 5-HT2 receptor agonist, and reversed by cyproheptadine, a selective 5-HT2 receptor antagonist. The potentiation of I(GABA) by 5-HT was irrespective to whether the I(5-HT) presents or not in a subset of neurons. The concentration-response curves for GABA before and after pretreatment with 5-HT manifested the same threshold value and similar EC50 (2.0 x 10(-5) and 1.9 x 10(-5) mol x L(-1), respectively) , while the maximal value of I(GABA) for the latter was 33.6% higher than that for the former. Intracellular dialysis with GDP-beta-S or H-7 abolished the potentiation of I(GABA) by 5-HT, while H-9 did not.

CONCLUSION5-HT can potentiate GABA-activated current via PKC-dependent phosphorylation of GABA(A) receptor following the activation of 5-HT2 receptor.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; pharmacology ; Animals ; Cyproheptadine ; pharmacology ; Female ; Ganglia, Spinal ; cytology ; physiology ; Male ; Membrane Potentials ; drug effects ; Neurons ; physiology ; Patch-Clamp Techniques ; Protein Kinase C ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley ; Receptors, Serotonin, 5-HT2 ; Serotonin ; analogs & derivatives ; pharmacology ; Serotonin 5-HT2 Receptor Agonists ; Serotonin 5-HT2 Receptor Antagonists ; Signal Transduction ; gamma-Aminobutyric Acid ; pharmacology

BACKGROUNDZengshengping (ZSP) tablets had inhibitory effects on oral precancerous lesions by reducing the incidence of oral cancer. However, the severe liver toxicity caused by systemic administration of ZSP limits the long-term use of this anti-cancer drug. The purpose of this study was to evaluate the tumor inhibitory effects due to the topical application of extracts from ZSP, a Chinese herbal drug, on 7, 12-dimethlbenz(a)anthracene (DMBA) induced oral tumors in hamsters. The study also investigated the anti-cancer mechanisms of the ZSP extracts on oral carcinogenesis.

METHODSDMBA (0.5%) was applied topically to the buccal pouches of Syrian golden hamsters (6 - 8 weeks old) three times per week for six weeks in order to induce the development of oral tumors. Different fractions of ZSP were either applied topically to the oral tumor lesions or fed orally at varying dosages to animals with oral tumors for 18 weeks. Tumor volume was measured by histopathological examination. Tumor cell proliferation was evaluated by counting BrdU labeled cells and by Western blotting for mitogen-activated protein kinase (MAPK) protein levels. The protein levels of apoptosis marker Caspase-3 and regulator Bcl-2 protein were also measured by Western blotting.

RESULTSTopical application of DMBA to the left pouch of hamsters induced oral tumor formation. Animals treated with DMBA showed a loss in body weight while animals treated with ZSP maintained normal body weights. Both the ZSP n-butanol fraction and water fraction significantly reduced tumor volume by 32.6% (P < 0.01) and 22.9% (P < 0.01) respectively. Topical application of ZSP also markedly decreased the BrdU-positive cell numbers in oral tumor lesions and reduced the expression level of MAPK. In addition, ZSP promoted tumor cell apoptosis by increasing Caspase-3 expression but decreasing Bcl-2 protein production.

CONCLUSIONThe n-butanol and water fractions of ZSP are effective at inhibiting tumor cell proliferation and stimulating apoptosis in oral cancer suggesting that these fractions have chemopreventive effects on DMBA induced oral carcinogenesis.

9,10-Dimethyl-1,2-benzanthracene ; toxicity ; Animals ; Antineoplastic Agents ; therapeutic use ; Cell Transformation, Neoplastic ; drug effects ; Cricetinae ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mesocricetus ; Mouth Neoplasms ; chemically induced ; drug therapy ; prevention & control

BACKGROUNDIn recent years, transfer of the spinal accessory nerve to suprascapular nerve has become a routine procedure for restoration of shoulder abduction. However, the operation via the traditional supraclavicular anterior approach often leads to partial denervation of the trapezius muscle. The purpose of the study was to introduce transfer of the spinal accessory nerve through dorsal approach, using distal branch of the spinal accessory nerve, to repair the suprascapular nerve for restoration of shoulder abduction, and to observe its therapeutic effect.

METHODSFrom January to October 2003, a total of 11 patients with a brachial plexus injury and an intact or nearly intact spinal accessory nerve were treated by transferring the spinal accessory nerve to the suprascapular nerve through dorsal approach. The patients were followed up for 18 to 26 months [mean (23.5 +/- 5.2) months] to evaluate their shoulder abduction and function of the trapezius muscle. The outcomes were compared with those of 26 patients treated with traditional anterior approach. And the data were analyzed by Student's t test using SPSS 10.5.

RESULTSIn the 11 patients, the spinal accessory nerves were transferred to the suprascapular nerve through the dorsal approach successfully. Intact function of the upper trapezius was achieved in all of them. In the patients, the location of the two nerves was relatively stable at the level of superior margin of the scapula, the mean distance between them was (4.2 +/- 1.4) cm, both the nerves could be easily dissected and end-to-end anastomosed without any tension. During the follow-up, the first electrophysiological sign of recovery of the infraspinatus appeared at (6.8 +/- 2.7) months and the first sign of restoration of the shoulder abduction at (7.6 +/- 2.9) months after the operation, which were earlier than that after the traditional operation [(8.7 +/- 2.4) months and (9.9 +/- 2.8) months, respectively; P < 0.05]. The postoperative shoulder abduction was 62.8 degrees +/- 12.6 degrees after transfer of the spinal accessory nerve, better than that after the traditional (51.6 degrees +/- 15.7 degrees). All the 11 patients could extend and externally rotate the shoulder almost normally.

CONCLUSIONSThe accessory nerve transfer through dorsal approach is a safe and reliable procedure for the treatment of brachial plexus injury. Its postoperative effect is confirmed, which is better than that of the traditional operation.

Accessory Nerve ; surgery ; Adolescent ; Adult ; Brachial Plexus ; injuries ; Humans ; Male ; Nerve Transfer ; methods ; Shoulder Joint ; innervation ; physiology

OBJECTIVETo investigate the effect of bone mesenchymal stem cell (BMSC) transplantation on repair of glomerular podocytes and on the Nephrin expression in rats with puromycin aminonucleoside (PAN) -induced nephrosis.

METHODSForty-five Sprague-Dawley rats were randomly divided into three groups (n=15 each): a nephrosis model group that received a single intraperitoneal injection of PAN (0.15 mg/g); a BMSC transplantation group that received a single intraperitoneal injection of PAN (0.15 mg/g) followed by BMSC transfusion; a control group that received a single intraperitoneal injection of normal saline. Ten days after injection, the rats were sacrificed. The 24 hrs urinary protein content and serum albumin and cholesterol levels were measured 24 hrs before sacrifice. Changes of glomerular podocytes were observed under an electron microscope. Brdu labeled positive cells in kidneys were measured by immunohistochemical technology. RT-PCR and Western blot were used to assess the expression of mRNA and protein of Nephrin.

RESULTSIn the nephrosis model group, urinary protein and blood cholesterol contents increased, plasma albumin content decreased compared with those in the control group. Extensive fusion of podocyte foot processes was observed in the nephrosis model group. The BMSC transplantation group had decreased urinary protein and blood cholesterol contents and increased plasma albumin content compared with the nephrosis model group. Fusion of podocyte foot processes was also improved. Brdu labeled positive cells were seen in kidneys in the BMSC transplantation group, but not in the nephrosis model and the control groups. Nephrin mRNA and protein expression decreased significantly in the nephrosis model group compared with that in the control group. The BMSC transplantation group had increased Nephrin mRNA and protein expression compared with the nephrosis model group.

CONCLUSIONSBMSCs can repair glomerular podocytes in PAN-induced nephrosis rats, and the changes of Nephrin expression may be involved in the process.

Animals ; Bromodeoxyuridine ; metabolism ; Kidney ; pathology ; Male ; Membrane Proteins ; genetics ; Mesenchymal Stem Cell Transplantation ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy ; Podocytes ; pathology ; Puromycin Aminonucleoside ; toxicity ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThe pulmonary toxicity of water extract of Siegesbeckia pubescens has been studied an mice following subchronic oral administration route.

METHODMice were randomly grouped and administered with the water extract of S. pubescens at dosages of 3.0, 1.0, 0.3 g x kg(-1) and saline respectively. 10 mice of each group were sacrificed on the day of 7 th, 14 th, 21 th, and 2 weeks after stopping administration, histological changes of the lung were examined.

RESULTThe water extract of S. pubescens at dosage of 3.0 g x kg(-1) increased the lung index on the of day 14 th and 21 th, significant histopathological damages were observed. The histopathological changes were disappeared after stopping administration for 2 weeks.

CONCLUSIONThe water extract of S. pubescens has a pulmonary toxic effect on mice, and the toxic effect is reversible.

Administration, Oral ; Animals ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Female ; Lung ; drug effects ; Male ; Mice ; Random Allocation

AIM:To detect the myeloid-derived suppressor cells(MDSCs )in peripheral blood from the pa-tients with Parkinson disease(PD)and its clinical significance.METHODS:The patients(n=80)diagnosed PD from January 2016 to March 2017 in our hospital and 20 healthy volunteers were selected as the subjects.According to the Hoehn-Yahr staging,80 PD patients were staged,of whom 22 were Ⅰ,24 were Ⅱ,20 were Ⅲ,14 were Ⅳ,and 0 was Ⅴ. Peripheral blood(5 mL)samples from the patients with PD and the healthy volunteers were collected and the mononuclear cells were isolated.The levels of CD14 +CD11b+cells and CD14 -CD11b+cells in the peripheral blood were detected by flow cytometry.The two populations of the cells were sorted by magnetic beads.The mRNA levels of arginase 1(ARG1),interleukin-10(IL-10)and cyclooxygenase 2(COX-2)were detected by qPCR.The expression of surface membrane pro-teins CD14 and CD11b,and immunosuppressive factors ARG1,IL-10 and COX-2 was determined by Western blot and ELISA.RESULTS:No significant change of CD14 +CD11b+cells between the patients with PD and normal controls was observed,but the cells with CD14 -CD11b+increased significantly in the patients with PD compared with the control peo-ple(P<0.05).The CD14 -CD11b+cells in peripheral blood of the patients were related to the stage of Hoehn-Yahr.The CD14-CD11b+and CD14+CD11b+cells showed high levels of IL-10 and COX-2,and the high level of ARG1 was only expressed in the CD14 -CD11b+cells.The expression of ARG1 in the CD14 -CD11b+population from PD patients was significantly different from that of CD14+CD11b+population and normal subjects(P<0.05).CONCLUSION:The CD14-CD11b+cells and ARG1 expression level in peripheral blood of the PD patients can be used to evaluate the patho -genesis and staging.Immunosuppression may play an important role in the pathogenesis and development of PD.