A lactoferrin-containing PEGylated liposome system (Lf-PLS) was developed and tested in vitro as a hepatoma-targeting drug delivery system. PEGylated liposomes (PLS) were successfully prepared using the thin film hydration method with peglipid post insertion. Lf was covalently conjugated onto the carboxyl terminal of DSPE-PEG2000-COOH on liposomes. Coumarin-6 was used to trace Lf-PLS with fluorescence. The cellular uptake of this system was carried out in asialoglycoprotein receptor (ASGPR) positive HepG2 cells via confocal microscopy and flow cytometry. The Lf-PLS liposome was observed as spherical or oval vesicles with the particle size around 130 nm, zeta potential about -30 mV and encapsulation efficiency more than 80%. The confocal microscopy images and flow cytometry data demonstrated that Lf-PLS resulted in significantly higher cell association by ASGPR positive HepG2 cells compared to PLS. The association between Lf-PLS and cells were dependent on the concentration, time and temperature, which was inhibited by pre-incubation with excessive free Lf. The results suggest that Lf-PLS has a good targeting effect on HepG2 cells in vitro. The targeting mechanism may be related to the specific binding of Lf and ASGPR on HepG2 cells, which guides Lf-PLS to the cell surface to induce an active endocytosis process. All these results demonstrated that Lf-PLS might be a potential drug delivery system in targeting hepatocellular carcinoma, which deserves more research on its targeting ability, antitumor efficiency, and metabolism in vivo for treatment of hepatomacellular carcinoma.
Asialoglycoprotein Receptor ; metabolism ; Carcinoma, Hepatocellular ; pathology ; Coumarins ; Drug Delivery Systems ; Endocytosis ; Hep G2 Cells ; drug effects ; Humans ; Lactoferrin ; pharmacology ; Liposomes ; Liver Neoplasms ; pathology ; Particle Size ; Phosphatidylethanolamines ; Polyethylene Glycols ; Thiazoles

Objective To evaluate and analyse the effectiveness of surgical treatment for fibrous dysplasia in proximal femur with severe varus deformity.Methods A retrospective study was performed in 21 patients (24 femora)of fibrous dysplasia who were treated in our hospital between August 2000 and May 2009.All patients had severe femoral varus deformity.The four-step procedures were performed orderlv as valgus osteotomy,lesion curettage,impacting of massive bone allograft,and fixation by femoral intramedullary nail.There were 6 patients with monostotic disease,15 with polyostotic diseases,including 12 males and 9 females with a mean age of 22.7 years(range,14-39 years).The average neck-shaft angle and femoral varus angle was 75°(range 55°-105°)and 30°(range,18°-45°),respectively.The average length of thigh shortened 3.4 cm(range,2.0-4.5 cm)compared with the contralateral thigh.Results All of the 21 patients were followed up from 21 months to 7 years with an average period of 3 years and 4 months.There were 30 osteotomy sites in 24 femurs,28 osteotomy sites showed bone union after 3-6 months from surgery.Two distal location of the double-level osteotomy showed nonunion,which received bone grafting again and got bone union after 3 months finally.The femoral mechanical alignments of the 21 patients had been recriticd.The average neck-shaft angle was 118°(range,95°-135°)postoperatively,the femoral varus angle disappeared.The average extremity lengthening was 2.8 cm(range,1.8-3.6 cm)postoperatively.There were no infection,recurrent fracture and progression of deformity.The visual analogue scales(VAS)score of 17 patients decreased to zero postoperatively from 7-10 preoperatively,and that of the other 4 patients decreased to 3-4 postoperatively from 8-10 preoperatively.The result of Harris hip functional score was excellent in 12 cases,good in 6,and fair in 3.Conclusion The valgus osteotomy can rectify varus deformity effectively.The reconstract nail of the fumer can support the stability of femur.Impacting of massive bone allograft can improve the capacity of the femur.

ObjectiveTo compare the neonatal outcomes between infants of assisted conception (AC) and natural conception (NC). Methods A prospective case-control study was conducted.Eligible mothers were invited to this study at 28 weeks of gestation and were followed up to delivery.All newborns were examined by pediatricians right after birth.Data including gestational age,birth weight,admission to neonatal intensive care unit (NICU),and presence of any major malformation were collected.Statistical analysis was performed with SPSS 13.0.Difference between two groups was compared by x2 test. ResultsSix hundred and seventy-three newborns were enrolled into this study,including 325 in AC group and 348 in NC group.There were no differences between the demographic characteristics of the two groups,including maternal age,parents' education,family income and proportion of primipara (P＞0.05).The incidences of twins (32.62％ vs 3.45％,x2 =98.88),preterm birth (30.46％ vs 7.76％,x2=56.92),low birth weight infant (32.31％ vs 8.91％,x2 =57.07)and admission to neonatal intensive care unit (27.38％ vs 8.91％,x2 =39.16) and proportion of cesarean birth (62.77％ vs 21.55％,x2=117.64) in AC group were significantly higher than in NC group (all P＜0.01),except for the incidence of birth defect (4.62％ vs 2.59％,x2=2.01,P＞ 0.05).However,no significant differences was found in the above items when only singletons were compared between the two groups (P＞0.05),except for the cesarean section rate (61.18％ vs 22.03％,x2 =100.93,P＜0.01).There were no difference in any of the above neonatal outcomes when different assisted reproductive technologies applied were compared(in vitro fertilization-embryo transfer,intracytoplasmic sperm injection and frozen embryo transfer) (P＞0.05).Conclusions Neonatal outcome after assisted conception is a bit worse than natural conception,which might mainly due to the large proportion of multiple pregnancy after assisted conception.In order to improve neonatal outcome after assisted conception,the number of embryos transferred should be limited.

HepG2 ceils were treated with various concentrations of tumor necrosis factor-or (TNF-α) for 24 hours. RT-PCR and Western blot were used to measure protein tyrosine phosphatase-1 B(PTP-1B)expression, and luciferase reporter assay was used to detect NF-kB activity. The results showed that treatment of HepG2 cells with TNF-α for 24 hours led to upregulation of PTP-1B and NF-kB activity in a dose-dependent manner. Inhibition of NF-kB by PI)TC significantly attenuated TNF-α-induced PTP-IB expression in HepG2 cells. Thus, the transactivation of NF-kB seems to play an important role in the expression of PTP-1B in HepG2 cells induced by TNF-α.

Cranial Fossa, Anterior ; Humans ; Liposarcoma, Myxoid ; diagnostic imaging ; metabolism ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; S100 Proteins ; metabolism ; Skull Base Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Tomography, X-Ray Computed

Objective To investigate the value of MR diffusion weighted imaging (DWI) in the pathological grading of glioma. Methods Thirty patients who were confirmed pathologically with glioma were examined by MR DWI. The values of apparent diffusion coefficient (ADC) and relative ADC (rADC) were measured. All those values were analyzed statistically. Results The 30 glioma patients were divided into lower-grade and higher-grade glioma groups. The lower-grade glioma group included 14 cases (1 case of WHO Ⅰ, tricholeukocyte astrocytoma; 13 cases of WHO Ⅱ, 11 astrocytomas with 1 case of post-operative reenrrence, 1 ependymoma, 1 oligodendroglioma); the higher-grade group included 16 cases (! I cases of WHOm, anaplastic astrocytoma with 1 case of post-operative recurrence; 5 cases of WHO Ⅳ, 4 multiforme glioblastomas, 1 ependymoma). The ADC and rADC values of lower-grade gliomas were (1.36±0.16)×10-3 mm2/s and 1.76±0.23, and the values of higher-grade glioma were (1.08±0.10)×10-3 mm2/s and 1.36±0.16, respectively. The values of ADC and rADC were significantly different between the 2 groups (P<0.05). The accuracy rates were 86.7% and 83.3% when the thresholds of ADC and rADC were 1.20×10-3 mm2/s and 1.53. Conclusion The values of ADC and rADC have high accuracy for pathological grading of the glioma.

Objective To investigate the value of perfusion-weighted magnetic resonanceimaging (MRI) in the grading ofgliomas. Methods Thirty patients with pathologically confirmedglioma underwent perfusion-weighted MRI. Of the 30 glioma patients, 14 patients had low-grade and 16had high-grade gliomas. The low-grade glioma patients included 1 with WHO grade Ⅰ glioma (pilocyticastroeytoma) and 13 with WHO grade Ⅱ gliomas; in the WHO grade Ⅱ patients, 11 had astrocytomaincluding I with postoperative recurrence, 1 with epeudymoma, and 1 with oligodendroglioma. Of the 16patients with high-grade gliomas, 11 had WHO grade Ⅲ gliomas identified as anaplastic astrocytoma(including 1 with postoperative recurrence) and 5 had WHO Ⅳ gliomas (including 4 with glioblastomamultiforme and 1 with ependymoma). The relative cerebral blood volume (rCBV), rMSD and relativemean transit time (rMTT) were measured and statistically analyzed in these patients. Results TherCBV, rMSD and rMTT were 1.99±1.0, 1.83±0.78, and 1.10±0.08 in the patients with low-grade gliomas,as compared to those in the high-grade giioma patients of 4.95±2.04, 3.59±1.13, and 1.03±0.61,respectively, showing significant differences in the rCBV and rMSD between the two groups (P<0.05).The diagnostic accuracy of rCBV and rMSD was 83.3% and 80% for the gliomas using their upper limits(2.99 and 2.61, respectively) as the diagnostic thresholds. Conclusion The values ofrCBV and rMSDhave high accuracy in the grading of brain gliomas.

In order to investigate the mechanism of acute lymphoblastic leukemic cell malignant proliferation, the expressions of hMSH2 mRNA and mutation P53 (mtP53) protein in bone marrow cells of de novo acute lymphoblastic leukemia (ALL) were determined by in situ hybridization and immunocytochemical methods. The results showed the that percentage of positive cell with hMSH2 mRNA expression was (32.88 +/- 11.46)% in the de novo ALL group and (64.22 +/- 8.51)% in the control group. The percentage of positive cell with mtP53 protein expression was (29.25 +/- 9.45)% in the de novo ALL group, and (12.63 +/- 6.66)% in the control group. There was a significant negative correlation between the positive percentages of hMSH2 mRNA expression and mtP53 protein expression (r = -0.45, P < 0.05). It is concluded that defective MSH2 mRNA expression plays an important role in the pathogenesis of acute lymphoblastic leukemia, mtP53 protein mutation plays an important role in the development of acute lymphoblastic leukemia, the hMSH2 mRNA defect can lead to accumulation of the mutant P53 protein in acute lymphoblastic leukemia, and both jointly promote the pathogenesis of ALL.
Adult ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Male ; MutS Homolog 2 Protein ; genetics ; Mutant Proteins ; biosynthesis ; Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVEA new pattern recognition method suitable for traditional Chinese patent medicine was established in this paper, which is named as the Dual index grade sequence pattern recognition.

METHODIn this method the quality gradation was defined mathematically relying on normal distribution. By this way samples can be clustered and classified depending on which quality gradation is wanted, and the grading samples quantitatively relative to quality can be performed simultaneously. Especially, the redundant information with respect to pattern recognition hiding in dual index sequences of samples can be removed effectively by applying the good grade sequences, which make the pattern recognition results accurate excellently. This approach possesses the advantages of both supervised classification and unsupervised cluster methods. Samples can be clustered and classified at the same time without any standard samples, and the operation is accomplished based on the good grade similar sequences themselves being as the classifying marks. Moreover, the subclasses in each class can be identified more subtly.

RESULTThe infrared fingerprint spectra of extracts of 27 kinds of Mingmu Dihuangwan pills and Zhibo Dihuangwan pills samples extracted with ethanol were analyzed with the method proposed in this paper. The results showed that these pills can be classified in their subclasses clearly, respectively.

CONCLUSIONThe Dual index grade sequence pattern recognition is a new and effective one for identifying complex biological products made from complex herbal medicines.

Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; classification ; Ethanol ; chemistry ; Pattern Recognition, Automated ; methods ; Quality Control ; Reproducibility of Results ; Spectrophotometry, Infrared

AIMTo observe the role of melatonin receptor and GABAA receptor in sleeping time prolonged by melatonin in mice.

METHODSThe absence of the righting reflex was considered as the sleep onset and the duration of the loss of the righting reflex was recorded as the sleeping time. The effects of receptor agonist and antagonist on hypnotic activity of melatonin were studied in the paper.

RESULTSPrazosin hydrochloride, the blocker of melatonin 3 receptor, didn't affect the sleeping time prolonged by melatonin in mice. GABA, the endogenous agonist of GABA receptor, significantly potentiated the hypnotic activity of melatonin. When picrotoxin, the ligand of picrotoxin site on GABAA receptor, used together with melatonin, it significantly antagonized the sleeping time prolonged by melatonin, however, bicuculline, the specific antagonist of GABA binding site in GABAA receptor, didn't affect the hypnotic activity of melatonin in mice.

CONCLUSIONMelatonin does not exhibit its potentiation sleeping time in mice through melatonin 3 receptor. Hypnotic activity of melatonin may be mediated through picrotoxin site on GABAA receptor.

Animals ; Bicuculline ; pharmacology ; Male ; Melatonin ; physiology ; Mice ; Mice, Inbred Strains ; Picrotoxin ; pharmacology ; Prazosin ; pharmacology ; Receptors, GABA-A ; physiology ; Receptors, Melatonin ; physiology ; Sleep ; physiology