OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

OBJECTIVEOsteopetrosis is a rare genetic disorder and the malignant infantile osteopetrosis (MIOP) is the worst subtype of this disease. Seventy percent of patients die in six years of life without proper treatment. Hematopoietic stem cell transplantation (HSCT) offers the only chance of cure for MIOP.

METHODRetrospective analysis was performed on 8 patients with MIOP who underwent HSCT in Beijing Children's Hospital during the period from 2006 to 2011.

RESULTEight cases (4 male and 4 female, mean age at HSCT 13.5 months) were diagnosed as malignant infantile osteopetrosis. Conditioning regimen included fludarabine, busulfan and cyclophosphamide. All patients received cyclosporin for prophylaxis of graft vs. host disease (GvHD). A UMD recipient underwent CD34(+) cell selection. ATG/ALG, mycophenolate mofetil (MMF) and methotrexate (MTX) used for recipients with unrelated cord donor (2) and recipients with haplo-identical donors (5). Average time for neutrophil engraftment was 15.7 day (9 - 36), platelet engraftment was 43.3 day (10 - 68). The patients were followed up from 47 days to 5 years, 1 patient died of post-transplant complications. Seven cases presented better in clinical manifestation. Acute GvHD I°-II° was observed in 6 patients, III°-IV° in 2 patients. It was controlled by anti-GvHD therapy.

CONCLUSIONNon-allogenic stem cell transplantation treatment of infantile MIOP showed high survival rate and restoration of hematopoiesis in haploid transplant patients, therefore, non-allogenic HSCT may be an option to treat MIOP in children.

Bone Marrow Transplantation ; adverse effects ; methods ; Child, Preschool ; Female ; Fetal Blood ; cytology ; Follow-Up Studies ; Genetic Predisposition to Disease ; Graft vs Host Disease ; drug therapy ; epidemiology ; prevention & control ; Haploidy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Infant ; Male ; Osteopetrosis ; mortality ; therapy ; Retrospective Studies ; Survival Analysis ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo investigate the intervention effect of Danggui Shaoyao San on rats with cirrhotic ascites, and discuss the effect of arginine vasopressin (AVP) on cirrhotic ascites.

METHODMale SD rats were randomly divided into the control group, the model group, Danggui Shaoyao San low, middle and high dose groups. The cirrhotic ascites rat model was established by CCl4 combined with phenobarbital. Their urines were collected at 24 h to observe urine excretion of each group. Filter papers were used to determine the amount of ascites. The levels of serum alanine aminotransferasa (ALT) , aspartate aminotransferase (AST) were detected by the automatic biochemistry analyzer. Plasma prothrombin time (PT) was evaluated by the blood coagulation analyzer. The concentration of AVP in plasma was detected by enzyme-linked immunosorbent assay (ELISA). Pathological changes in livers were observed by HE staining.

RESULTCompared with the model group, the Danggui Shaoyao San group showed significant improvement in live indexes, with notable decrease in serum ALT and AST and the time of PT, improvement in liver pathological changes. Simultaneously, the amount of ascites decreased to varying degrees, with notable increase in urine in 24 h and decrease in AVP concentration in plasma.

CONCLUSIONDanggui Shaoyao San can notably improve liver functions of rats with cirrhotic ascites, reduce the generation of ascites and delay the progress of liver pathological changes. Its mechanism may be related to AVP.

Animals ; Arginine Vasopressin ; blood ; Ascites ; blood ; complications ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Liver ; drug effects ; enzymology ; metabolism ; physiopathology ; Liver Cirrhosis ; complications ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effects of pre-acupuncture and immediate acupuncture on kidney function and oxygen free radical metabolism in rats with simulated weightlessness.

METHODSTwenty male clean-grade Wister rats were randomly divided into a normal control group, a model group, a pre-acupuncture group and an immediate acupuncture group, 5 rats in each one. The rats in the normal control group did not receive any treatment but free activities for 4 weeks. The rats in the rest groups received 4-week tail suspension to establish the model of simulated weightlessness. One week before the tail suspension, the rats in the pre-acupuncture group were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once a day for 7 days. The rats in the immediate acupuncture group received tail suspension and acupuncture at the same time; during the tail suspension, the electroacupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once every other day for 14 days. The colorimetric method was used to measure the content of blood urea nitrogen (BUN) in serum as well as activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) and content of malonaldehyde (MDA) in renal tissue in each group.

RESULTSCompared with the normal control group, the content of BUN in the model group was increased significantly (P<0.01), the activity of SOD and GSH-PX in nephridial tissue was significantly reduced (both P<0.01), and the content of MDA was increased significantly (P<0.05). Compared with the model group, the content of BUN in the pre-acupuncture group and immediate acupuncture group was significantly reduced (P<0.01, P<0.05), the activity of GSH-PX in the pre-acupuncture group was obviously increased (P<0.05) and the content of MDA in the immediate acupuncture group was increased significantly (P<0.01). Compared with the immediate acupuncture group, the content of MDA in the pre-acupuncture group was lower (P<0.01).

CONCLUSIONThe pre-acupuncture and immediate acupuncture both have the capacity to improve the kidney function and anti-oxygen free radical injury in rats with simulated weightlessness, however, the capacity to increase the protection ability of the kidney and eliminate free radical in the pre-acupuncture group is superior to that in the immediate acupuncture group, which is likely to be related with improving antioxidant ability of kidney.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Antioxidants ; metabolism ; Humans ; Kidney ; metabolism ; Kidney Diseases ; metabolism ; physiopathology ; therapy ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Space Flight ; Superoxide Dismutase ; metabolism ; Weightlessness ; adverse effects

This study was aimed to establish the technique of interphase fluorescence in situ hybridization (I-FISH) used on smear of bone marrow directly, and to develop a new method for detection of the molecular cytogenetics in multiple myeloma (MM). After a series of treatment, fixation and digestion of the bone marrow smear as the carrier, the chromosome 8 centromere probe were used in I-FISH for molecular cytogenetics detection. At the same time, differences were compared in the results between the new method and the conventional I-FISH. The results showed that there was no statistically significant difference of proportion of various signals in non-hematologic malignancies when detected with the two methods (p>0.05). In bone marrow smear I-FISH, 8 out of 19 cases (42.1%) had abnormality of chromosome 8, including 5 cases with -8 (26.3%) and 3 cases with +8 (15.8%). It is concluded that the I-FISH on smear of bone marrow is characterized by convenience, economy and accuracy. Therefore, it can be used for research of molecular cytogenetics in MM.
Adult ; Aged ; Bone Marrow ; pathology ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

Diabetic retinopathy(DR)is a prevalent microvascular complication of diabetes and the leading cause of blindness and severe visual impairment in adults.The high levels of glucose trigger multiple intracellular oxidative stress pathways,such as POLDIP2,resulting in excessive reactive oxygen species(ROS)pro-duction and increased expression of vascular cell adhesion molecule-1(VCAM-1),hypoxia-inducible factor 1α(HIF-1α),and vascular endothelial growth factor(VEGF),causing microvascular dysfunction.Dihydromyricetin(DMY)is a natural flavonoid small molecule antioxidant.However,it exhibits poor solubility in physiological environments,has a short half-life in vivo,and has low oral bioavailability.In this study,we present,for the first time,the synthesis of ultra-small Fe-DMY nano-coordinated polymer particles(Fe-DMY NCPs),formed by combining DMY with low-toxicity iron ions.In vitro and in vivo experiments confirm that Fe-DMY NCPs alleviate oxidative stress-induced damage to vascular endo-thelial cells by high glucose,scavenge excess ROS,and improve pathological features of DR,such as retinal vascular leakage and neovascularization.Mechanistic validation indicates that Fe-DMY NCPs can inhibit the activation of the Poldip2-Nox4-H2O2 signaling pathway and downregulate vital vascular function indicators such as VCAM-1,HIF-1α,and VEGF.These findings suggest that Fe-DMY NCPs could serve as a safe and effective antioxidant and microangio-protective agent,with the potential as a novel multimeric drug for DR therapy.

Aim: To investigate the alleviating effect of NMDA receptor blocking on learning and memory impairment induced by gp120 in rats and its mechanism. Methods: (1 ) Thirty-two SD rats were randomly divided into control group, sham operation group, gpl20 group, and gp120 + Memantine group. Except for the control group, the other groups underwent a bilateral hippocampal injection to establish the model of learning and memory impairment in rats. Memantine (10 mg • kg