OBJECTIVETo study the effect of Guishen Pill (GSP) on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve (DOR).

METHODSTotally 40 female C57BL/6J mice were randomly divided into 4 groups, the normal control group, the model group, the GSP group, and the dehydroepiandrosterone (DHEA) group, 10 in each group. Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG), and prostaglandin F2α (PGF2α) were sequentially administrated to produce superovulation. The DOR model was established by exposing to ozone inhalation. Mice in the GSP group were intragastrically administered with GSP at 0.3 mL. Those in the DHEA group were intragastrically administered with DHEA at 0.3 mL. Equal volume of normal saline was intragastrically administered to mice in the normal control group and the model group. All mice wer treated for 21 days. Serum levels of estrogen (E2), progestogen (P), and anti-Müllerian hormone (AMH) were measured by ELISA. Changes of Oct-4, anti-AMH, and early growth response gene-1 (Egr-1) mRNA in ovaries were dtected by Real-time PCR.

RESULTSCompared with the model group, serum levels of E2, P, and AMH, as well as contents of estrogen receptor (ER), progestogen receptor (PR), MVH, and Oct-4 mRNA significantly increased in the GSP group and the DHEA group (P < 0.05).

CONCLUSIONGSP could improve expression levels of Oct-4, MVH, and Egr-1 mRNA in DOR mice and their ovarian function.

Animals ; Anti-Mullerian Hormone ; metabolism ; Dehydroepiandrosterone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Early Growth Response Protein 1 ; metabolism ; Estrogens ; Female ; Mice ; Mice, Inbred C57BL ; Octamer Transcription Factor-3 ; metabolism ; Ovarian Reserve ; Ovary ; Pregnancy ; Receptors, Estrogen ; metabolism ; Superovulation

Objective To study the neuroprotective effect of fucoidan against 6-OHDA-induced damage on MN9D cell line and preliminarily explore its mechanism.Methods The MN9D cells were treated with different concentrations of 6-OHDA(12.5,25,50,100 and 200 μmol/L)for 24 h and with 50 μmol/L 6-OHDA for different time courses(6,12,24 and 48 h)as well.Based on the above experiment,the best concentration(50 μmol/L)of 6-OHDA and time course(24 h)were selected and the damaged model of MN9D cell line was induced accordingly.The MN9D cells were pretreated with 0.01,0.1 and 1.0 mg/mL fucoidan for 1 h,and then co-treated with 50 μmol/L 6-OHDA for 24 h.The viability of MN9D cells was detected by MTT assay;the LDH leakage was detected by bioassay;the level of intracellular xidative stress was observed by 2',7'-dichlorodihydrofluorescein diacetate assay.Results MTT metabolic value decreased following the increase of 6-OHDA concentration or the extension of processing time.Treatment with 50 μmol/L 6-OHDA for 24 h induced a significant decrease of MTT metabolic rate and an increase of LDH leakage rate in the MN9D cells.Pretreatment with 0.1 and 1.0 mg/mL fucoidan inhibited the death of 6-OHDA-induced cells,increased the MTT metabolic value and decreased the LDH leakage.Cell morphological changes were consistent with the biochemical experiments.Treatment with 50 μmol/L 6-OHDA for 8 h could significantly increased the generation of reactive oxygen species induced by 6-OHDA,while pretreatment with 1.0 mg/mL fucoidan for 1 h showed antagonism on that.Conclusion Fucoidan can protect MN9D cells from the death induced by 6-OHDA and the underlying mechanism may be involved in the antioxidant capacity of fucoidan.

Objective:Tacrolimus prolonged-release(PR) formulation is a new once-daily formulation of the calcineurin inhibitor tacrolimus,which is currently used in adult liver or kidney transplant patients,and is also gradually widely used in children with nephrotic syndrome.The present study was undertaken to preliminarily investigate the pharmacokinetic characteristics of tacrolimus PR in pediatric nephrotic syndrome recipients.Methods:This single-center open-label prospective study was performed in pediatric nephrotic syndrome recipients.Pharmacokinetic samples were collected from eight pediatric subjects with nephrotic syndrome from Department of Pediatric Nephrology in Peking University First Hospital between June and August 2011.They followed administration of single oral doses of tacrolimus PR formulation at 0.02 mg/kg (n =2),0.05 mg/kg (n =2) and 0.10 mg/kg (n =4).Blood samples were taken before the dose and 1,2,4,6,8,10,12 and 24 h after drug intake.No other medicines or interacting food or drinks were taken during the study period.Blood concentrations were measured using an enzyme multiplied immunoassay technique.Pharmacokinetic analysis was performed using WinNolin Phoenix software Version 6.0 (Pharsight,Cary,NC,USA).Results:The pharmacokinetic data were best described by a non-compartment model.Pharmacokinetic parameters of tacrolimus PR formulation in the 3 ascending doses groups (0.02 mg/kg,0.05 mg/kg and 0.10 mg/kg) were as follows:the maxi mum drug concentrations (Cm=/D) were (1.7 ± 1.0) μg/L,(3.1 ± 1.9) μg/L,(8.0 ± 3.5) μg/L,respectively;Areas under the drug concentration-time curve (AUCo-∞/D) were (47.2 ± 47.1) h · μg/ L,(84.0 ± 13.1) h · μg/L,(175.6 ± 107.1) h · μg/L,respectively;Oral clearance rates were (0.8±0.9) L/(h·kg),(0.4±0.1) L/(h · kg),(1.9 ±1.3) L/(h · kg),respectively;Body weight normalized distribution volumes were (7.0 ± 3.4) L/kg,(12.4 ± 8.4) L/kg and (73.6 ± 68.6) L/kg,respectively.Both mean Cmax normalized level for the administered dose (Cmax/D) and mean AUC0-∞ normalized level for the administered dose (AUC0-∞/D) were higher in the 0.05 mg/kg dosage group than in the 0.02 and 0.10 mg/kg dosage group.There were two peaks in the drug concentrations in every dose group;a primary peak appeared at the end of about 2 h followed by a small secondary peak at h 12,which was more noticeable in the 0.10 mg/kg dose group than in the two lower dosages.Conclusion:The pharmacokinetic characteristics of tacrolimus PR formulation were initially explored in pediatric patients with nephritic syndrome.The data presented form a basis for subsequent larger scale studies on pharmacokinetics of tacrolimus PR formulation in nephritic syndrome children.

OBJECTIVETo compare between MLO-Y4 osteocyte and osteoblast to support osteoclast formation in co-culture system.

METHODSMLO-Y4 cells and murine osteoblast cells were co-cultured with bone marrow cells with or without vitamin D₃ presence.Bone marrow cells were as control group. Tartrat resistant acid phosphatase (TRAP)+ giant cells with three or more nuclei were counted and compared under a microscope at day 9.

RESULTSIn the absence of vitamin D₃, (1963.3 ± 93.1)/plate osteoclasts were observed when MLO-Y4 cells co-cultured with bone marrow cells in 24-well plate.While only (12.7 ± 5.5)/plate osteoclasts were found in the osteoblast group, and (6.0 ± 1.0)/plate in control group. The statistical difference occurs for any two groups (P < 0.05). Vitamin D₃ could significantly increase osteoclast formation in the three groups.

CONCLUSIONSOsteocytes could induce osteoclastogenesis without the presence of vitamin D₃ and vitamin D₃ could enhance the induction effects of MLO-Y4 and osteoblast cells.

Animals ; Cell Line ; Cholecalciferol ; chemistry ; Coculture Techniques ; Culture Media ; chemistry ; Mice ; Osteoblasts ; cytology ; Osteoclasts ; cytology ; Osteocytes ; cytology

The aim was to study the protective effects of amifostine (AMF) on normal hematopoietic stem/progenitor cells against the chemotherapeutic damage from etoposide (VP-16). The cord blood mononuclear cells (CBMNC), fresh and frozen peripheral blood stem cells (PBSC), and HL-60 cells were divided into AMF, AMF + VP-16, VP-16 and control groups, each group cell viability was determined by using trypan blue exclusion test, the CFU-GM culture was used to count cells, the apoptosis was detected by flow cytometry. The results showed that in CBMNC, fresh and frozen PBSC samples, cell viability and the number of CFU-GM in AMF + VP-16 group were all significantly higher than those in VP-16 group (P < 0.05); the CFU-GM incidence in AMF + VP-16 group was higher than that in VP-16 group, and the GFU-GM life in AMF + VP-16 group was also longer than that of latter, in CBMNC samples, the number of CFU-GM in AMF groups was higher than that in control group, but there was no statistical significance between the two groups (P > 0.05), in HL-60 cell apoptotic rate in AMF + VP-16 group was little higher than that in VP-16 group, but no statistical significance between these two groups (P > 0.05). It is concluded that AMF can significantly protect normal hematopoietic stem/progenitor cells against the damage from VP-16. Moreover, AMF does not affect cytotoxity of VP-16 on HL-60 cells, and can not stimulate the growth and differentiation of cord hematopoietic stem/progenitor cells directly.
Amifostine ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Etoposide ; pharmacology ; Flow Cytometry ; HL-60 Cells ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; Protective Agents ; pharmacology

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

Fucoidan is a natural polysaccharide extracted from brown seaweeds, with a wide variety of biological features, especially the anti-inflammatory and anti-oxidative effects. Studies indicated that the anti-inflammatory effect of fucoidan related to its capacity to interact with the selectin or scavenger receptor on the cell membrane. Fucoidan can also inhibit the synthesis and release of reactive oxygen species (ROS), as well as promote its clearance, showing the anti-oxidative activity.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; metabolism ; pharmacology ; Antioxidants ; isolation & purification ; pharmacology ; Polysaccharides ; isolation & purification ; metabolism ; pharmacology ; Reactive Oxygen Species ; metabolism ; Receptors, Scavenger ; metabolism ; Seaweed ; chemistry ; Selectins ; metabolism

AIMTo evaluate how solution viscosity affects the precorneal residence of five water-soluble polymers with different properties.

METHODSCaptive bubble technique was used, with the consecutive change of contact angle interpreted as an indication of desorption process, to study the residence of those polymers in vitro on freshly enucleated rabbit eyes under physiological conditions.

RESULTSCarbopol and sodium hyaluronate (HA), which adsorbed to isolated ocular surface more than 15 min, showed the optimum precorneal retentive capabilities. When the solution viscosity increased from 12 mPa.s to 50 mPa.s, the residence time of carbopol and HA were prolonged 10 min and 7 min, respectively, but that of sodium carboxymethylcellulose was not affected.

CONCLUSIONThe result suggested that higher viscosity is beneficial to improve the ocular residence time of bio-adhesive polymers.

Acrylic Resins ; Adhesiveness ; Animals ; Cornea ; drug effects ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Female ; Hyaluronic Acid ; pharmacokinetics ; pharmacology ; In Vitro Techniques ; Male ; Polyvinyls ; pharmacokinetics ; pharmacology ; Rabbits ; Solutions ; Viscosity

The aim of the study is to explore the effects of luteolin preconditioning on hepatic ischemia/reperfusion injury in rats and its mechanism, and investigate the effects of the change of heme oxygenase-1 (HO-1) activity on hepatic ischemia/reperfusion injury. Sprague-Dawley rats were divided into 5 groups randomly: control, model, luteolin, luteolin + zinc protoporphyrin (ZnPP, an inhibitor of HO-1) and hemin groups (n = 8 for each group). The rats in control, model and hemin groups received a standard chow daily. The rats in luteolin and luteolin + ZnPP groups received a chow supplemented with luteolin (200 mg/kg) daily. After 4 weeks, ZnPP (25 μmol/kg) and hemin (20 μmol/kg) were injected hypodermically 6 h before ischemia/reperfusion in luteolin + ZnPP and hemin groups, respectively. Portal vein and hepatic artery supplying the middle and left hepatic lobe were clamped with an atraumatic vascular clip for induction of partial hepatic ischemia in all rats except control group. After the 60 min of hepatic ischemia, a 60-minute reperfusion period was initiated by removal of the arterial clip. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected in serum, and the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum and liver were measured with assay kit. The expression of HO-1 protein and activity of HO-1 were examined in liver. The results showed that the luteolin and hemin pretreatment led to significant decreased levels of AST and ALT in serum, increased activity of SOD and decreased content of MDA in serum and liver compared with model group (P < 0.01). In addition, the expression of HO-1 protein and activity of HO-1 were elevated in luteolin and hemin groups (P < 0.01). ZnPP markedly increased the levels of AST and ALT in serum, and decreased the activities of SOD and HO-1, elevated MDA content in liver when compared with those in luteolin group (P < 0.01). Cytoplasmic vacuolation and swelling of hepatocytes were revealed in the model group after ischemia/reperfusion. Treatments with luteolin and hemin markedly relieved the liver structural changes. These results suggest that HO-1 protects rat liver from ischemia/reperfusion injury, and luteolin reduces the content of MDA and increases the activity of SOD and the expression of HO-1, which indicate that luteolin can elevate the antioxidation in rat liver, and thus protects rat liver from ischemia/reperfusion injury.
Animals ; Female ; Heme Oxygenase (Decyclizing) ; metabolism ; Ischemic Preconditioning ; methods ; Liver ; blood supply ; Luteolin ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

OBJECTIVEDevelopment and use of better medicine for children is a worldwide problem recently, especially in China. The current situation of drugs for children's renal diseases is far from well-understood now. This survey focused on drugs for pediatric renal diseases including immunosuppressants, corticosteroids, diuretics, anticoagulants, hypotensives and antilipemic agents.Information regarding the dosage, form, precaution, usage and administration in inserts was collected in this study.

METHODDrugs for pediatric renal diseases were selected according to the guidelines established by the Chinese Society of Pediatric Nephrology. The detailed information about the dosage, form of drugs was searched on the website of China-State Food and Drug (SFDA). The information of the precaution, usage and administration was obtained from the China Pharmaceutical Reference, the first edition.

RESULTIn this study, there were 5 categories of medicine including immunosuppressants, corticosteroids, diuretics, anticoagulants, hypotensives and antilipemic agents, and 89 kinds of medicine for renal diseases. Among these medicines, 65.2% were found not suitable for children in terms of drug dosage and form, 19.1% did not indicate the precaution, 51.7% did not indicate clearly the safety and effectiveness for children, and 56.2% lacked the detailed information about the usage and administration for children. There were only 4 kinds of these medicines which were studied via clinical trials in children population.

CONCLUSIONThere is a lack of drugs for children with renal diseases. Most of the time, the medicines used by doctors are not specially manufactured for children. The safety and efficacy of drugs that are currently used to treat pediatric renal diseases are not clear and definite.In addition, few clinical trials have been conducted for evaluation of drugs for pediatric renal diseases.In clinic, the situation of off-label drug treatment is very serious.

Adrenal Cortex Hormones ; administration & dosage ; therapeutic use ; Child ; Child, Preschool ; China ; Diuretics ; administration & dosage ; therapeutic use ; Dosage Forms ; Drug Approval ; Drug Labeling ; Drug Therapy ; standards ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Infant ; Kidney Diseases ; drug therapy ; Medication Therapy Management ; statistics & numerical data ; Off-Label Use ; statistics & numerical data ; Pediatrics