The latest findings of our laboratory showed that Angelica sinensis polysaccharide (ASP) showed a definite effect in regulating the aging of hematopoietic stem cells. Leukemia is a type of malignant hematopoietic tumor in hematopoietic stem cells. There have been no relevant reports about ASP's effect in regulating the aging of leukemia cells. In this study, human acute myeloid leukemia (AML) KG1alpha cell lines in logarithmic growth phase were taken as the study object, and were divided into the ASP group, the cytarabine (Ara-C) group, the ASP + Ara-C group and the control group. The groups were respectively treated with different concentration of ASP, Ara-C and ASP + Ara-C for different periods, with the aim to study the effect of ASP combined with Ara-C in regulating the aging of human acute myeloid leukemia KG1alpha cell lines and its relevant mechanism. The results showed that ASP, Ara-C and ASP + Ara-C could obviously inhibit KG1alpha cell proliferation in vitro, block the cells in G0/G1 phase. The cells showed the aging morphological feature. The percentage of positive stained aging cells was dramatically increased, and could significantly up-regulate the expression of aging-related proteins P16 and RB, which were more obvious in the ASP + Ara-C group. In conclusion, the aging mechanism of KG1alpha cell induced by ASP and Ara-C may be related to the regulation of the expression of aging-related proteins, suggesting that the combined administration of ASP and anticancer drugs plays a better role in the treatment of leukemia .
Aging ; drug effects ; genetics ; metabolism ; Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Polysaccharides ; pharmacology ; Retinoblastoma Protein ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo determine the safety and efficacy of local administration of lentivirus-mediated small interfering RNA (siRNA) targeting tumor necrosis factor-alpha (TNF-alpha) in murine air pouch model.

METHODSFrom May 2007 to April 2008 a siRNA targeting TNF-alpha and a missense siRNA were designed, and recombine lentivirus which coexpressed the green fluorescent protein (GFP) as a marker gene was constructed. Air pouches were established and stimulated by Ti-6Al-4V particles. Pouches were divided into 3 groups randomly. Lentivirus-mediated siRNA targeting TNF-alpha (TNF-alpha group) or lentivirus-mediated missense siRNA (MS group), or virus-free saline (control group) were injected into pouches respectively. Pouch membrane, peripheral blood, heart, liver, spleen, kidney, lung and brain were harvested at 28 d after transfection, and assayed for markers of inflammation using histological, molecular, immunological techniques and Xenogen in vivo imaging system (IVIS) 50 vivo bioluminescent assay system.

RESULTSXenogen IVIS 50 vivo image revealed strong expression of GFP localized in pouch areas and no expression in other parts of mice both in TNF-alpha group and MS group at 4 weeks after transfection, while no expression of GFP was found in control group. By RT-PCR and ELISA, the mRNA and protein levels of TNF-alpha in TNF-alpha group decreased by 81.6% and 82.6% respectively compared to control group (P < 0.01), and decreased by 78.9% and 84.0% respectively compared to MS group (P < 0.01), whereas TNF-alpha level in peripheral blood, heart, liver, spleen, kidney, lung and brain remained invariant (P > 0.05). Less inflammatory responses (thinner pouch membrane and decreased cellular infiltration) were observed in TNF-alpha group.

CONCLUSIONEfficient local delivery of lentivirus-mediated siRNA targeting TNF-alpha into modified murine air pouch can inhibit debris-induced inflammation effectively, with no systemic adverse effects.

Animals ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; genetics ; Inflammation ; therapy ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; genetics ; Random Allocation ; Transfection ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo build the physical model of four suturae which are related to the growth of maxilla in the three-dimensional finite-element model of maxillofacial bones.

METHODSA 16 years old volunteer with individual normal occlusion, good periodontium health condition and without diseases of temporomandibular joint was chosen to be the material of modeling. The three-dimensional finite-element model of the volunteer's maxillofacial bones was built using the CT scan and the finite-element modeling method. Finally we built the physical model of four suturae which were related to the growth of maxilla in the model of maxillofacial bones.

RESULTSThe model of maxillofacial bones with 86,575 nodes and 485,915 elements was generated. This model contained four suturae including sutura frontomaxillaris, sutura zygomaticomaxillaris, sutura temporozygomatica and sutura pterygopalatine.

CONCLUSIONA three-dimensional finite-element model of maxillofacial bones with good biological similarity was developed.

Bone and Bones ; Cranial Sutures ; Finite Element Analysis ; Head ; Humans ; Male ; Maxilla ; Skull ; Temporomandibular Joint ; Tomography, X-Ray Computed

OBJECTIVETo establish a mouse model of biliary obstruction.

METHODSSixty-four Balb/c mice were divided into experimental group and control group. Obstructive jaundice was induced in the mice in the experimental group by common bile duct ligation. The level of the common bile duct diameter, WBC, LYM MID, LYM%, MID% and ALT, AST, TBIL, DBIL, IBIL, ALP and CHOL were measured 12 h and 1, 2 ,3, 4, 5, and 7 days after the ligation. The morphological changes in the liver were also observed.

RESULTSThe level of common bile duct diameter, WBC, LYM, MID, LYM%, MID% and ALT, AST, TBIL, DBIL, ALP and CHOL all underwent changes with time following certain patterns.

CONCLUSIONThe jaundice manifestation of this model is similar to that of patients with biliary obstruction, and this model may provide a reliable model for studying the mechanism of obstructive jaundice.

Animals ; Cholestasis, Extrahepatic ; pathology ; Common Bile Duct ; pathology ; surgery ; Disease Models, Animal ; Female ; Ligation ; Liver ; pathology ; Mice ; Mice, Inbred BALB C

BACKGROUNDConnective tissue growth factor (CTGF) contributes greatly to renal tubulointerstitial fibrosis, which is the final event leading to end-stage renal failure. This study was designed to investigate the effects of CTGF antisense oligodeoxynucleotides (ODNs) on the expressions of plasminogen activator inhibitor-1 (PAI-1) and fibronectin in renal tubular cells induced by transforming growth factor beta1 (TGF-beta1) in addition to the role of CTGF in the accumulation and degradation of renal extracellular matrix (ECM).

METHODSA human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODNs were transfected into HKC cells. After HKC cells were stimulated with TGF-beta1 (5 microg/L), the mRNA levels of PAI-1 and fibronectin were measured by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 and fibronectin in the medium were determined by Western blot and ELISA, respectively.

RESULTSTGF-beta1 was found to induce tubular CTGF, PAI-1, and fibronectin mRNA expression. PAI-1 and fibronectin mRNA expression induced by TGF-beta1 was significantly inhibited by CTGF antisense ODNs. CTGF antisense ODNs also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 and fibronectin protein secreted into the medium.

CONCLUSIONSCTGF may play a crucial role in the accumulation and degradation of excessive ECM during tubulointerstitial fibrosis, and transfecting CTGF antisense ODNs may be an effective way to prevent renal fibrosis.

Cells, Cultured ; Connective Tissue Growth Factor ; Extracellular Matrix ; metabolism ; Fibronectins ; genetics ; secretion ; Humans ; Immediate-Early Proteins ; genetics ; physiology ; Intercellular Signaling Peptides and Proteins ; genetics ; physiology ; Kidney Tubules ; metabolism ; Oligonucleotides, Antisense ; pharmacology ; Plasminogen Activator Inhibitor 1 ; genetics ; secretion ; RNA, Messenger ; analysis ; Transfection ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1

OBJECTIVETo discuss the treatment of the scrotal vein malformation in teenagers and clinical efficacy.

METHODS32 cases with the local and diffuse scrotal vein malformation were retrospectively analyzed. 31 cases underwent local injection with 40% urea before resection. The urea was injected locally into tumor through multi-points within 30 seconds, 2-6 ml every time, one time a day. The injection was performed for 5-12 days. The treatment was refused in one case. The therapeutic effect and cosmetic result were recorded.

RESULTSThe tumors were removed radically in 28 cases including one operation in 25 cases and secondary operation in 3 cases. The patients were followed up for 1-3 years with no recurrence. Cosmetic result with bilaterally symmetric scrotum was satisfactory. The tumors in 3 severe cases were partially resected with improvement.

CONCLUSIONSCombined treatment with urea injection and surgical procedure can effectively treat the scrotal vein malformation with satisfactory result.

Adolescent ; Humans ; Injections, Intralesional ; Male ; Retrospective Studies ; Scrotum ; blood supply ; Urea ; administration & dosage ; therapeutic use ; Vascular Malformations ; therapy ; Young Adult

OBJECTIVETo explore the relationship between children's motor coordination and their physical growth to provide clues for clarifying the causal relationship between them and to effectively enhance the children's physical growth.

METHODTen kindergartens were selected randomly from district of Hongkou, Putuo and Pudong in Shanghai, and 1099 children were included in the analysis. The children's motor coordination was assessed using Developmental Coordination Disorder Questionnaire (DCDQ). The relationship between the children's motor coordination and their physical growth were analyzed by Structural Equation Model (SEM). The above relationship by different gender, age and BMI (overweight or not) were analyzed with multi-group SEM.

RESULTSOf 1099 subjects, 561 were boys (51%), and 538 were girls (49%). Of the total subjects, 354 (32.2%) were children aged 3-years old, 441 (40.1%) were children aged 4-years, 276 (25.2%) were children aged 5-years, and 28 were children (2.5%) aged 6-years old. The result of SEM showed that the children's gross motor, fine motor and general motor ability was correlated positively with children's physical growth (P < 0.001), their standardized estimation of structural coefficient were 0.40, 0.36, 0.37 respectively. The result of multi-group SEM showed that there was no significant difference in the SEM between different gender and ages. The fine motor ability was positively correlated with child physical growth (P < 0.001). There was no inter-group difference in the structural model (each P > 0.05). However, there was difference in the structural model between overweight and normal group, of them, only the structural coefficient of gross motor and physical growth were different between the two groups (t = 1.697, P < 0.05).

CONCLUSIONThere is a relationship between children's motor coordination and physical growth. The degree of association between the gross moter and physical growth in overweight children were different from the normal children. Further study on the factors influencing the children's physical growth is needed.

Child Development ; Child, Preschool ; China ; Female ; Humans ; Male ; Motor Skills ; Surveys and Questionnaires

OBJECTIVETo investigate the serotypes distribution and ampicillin resistance of Haemophilus influenzae isolates from children with respiratory infection in Hangzhou.

METHODSHaemophilus influenzae strains were identified with V factor and X factor tests. Serotypes were determined with the slide agglutination method. Nitrocefin test was used to detect beta-lactamase. The sensitivities of ampicillin to Haemophilus influenzae were determined with the Kirby-Bauer diffusion method and the E-test method.

RESULTSOne hundred and fifty-two Haemophilus influenzae isolates, 108 from boys and 44 from girls, were identified between December 2006 and July 2007. Of the 152 isolates, 148 (97.4%) were untypable, only 4 (2.6%) were typable, including type a, type d, type e and type f (n=1 each type). Haemophilus influenzae type b and c strain was not found. Thirty-four isolates (22.4%) were beta-lactamase-positive. One hundred and thirteen isolates (74.3%) were susceptible to ampicillin, while 34 isolates (22.4%) were resistant to ampicillin.

CONCLUSIONSUntypable strains were the most common in Haemophilus influenzae isolates from children with respiratory infection in Hangzhou. The isolates of Haemophilus influenzae kept susceptibity to ampicillin to a certain extent.

Adolescent ; Ampicillin Resistance ; Child ; Child, Preschool ; Female ; Haemophilus influenzae ; classification ; drug effects ; enzymology ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Respiratory Tract Infections ; microbiology ; Serotyping ; beta-Lactamases ; analysis

The methods to determine the total phenols, total saponins, and marker constituents salidroside, chlorogenic acid and 3, 4-dihydroxy-phenylethyl-β-D-glucopyranoside in the samples of Sargentodoxae Caulis were established to provide the evidence for the improvement and revision of the quality standard of the crude material recorded in the Chinese Pharmacopoeia (2015 Edition). The content of total phenols was determined by ultraviolet spectrophotometry, using gallic acid as a reference substance. The content of total saponins was determined by ultraviolet spectrophotometry, using 3-O-[β-D-xylopyranosyl-(1-2)-O-β-D-glucuronopyranosyl]-28-O-[β-D-glucopyranosyl] asiatic acid as a reference substance. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside were detected by HPLC. The linear ranges were 1.01-7.04 mg x L(-1) for total phenols, 37.7-201 μg for total saponins, 0.025 8-1.55 μg for salidroside, 0.076 2-5.44 μg for chlorogenic acid, and 0.064 9-3.47 μg for 3,4-dihydroxy-phenylethyl-βP-D-glucopyranoside, respectively. Their average recoveries were 99.12%, 99.11% 105.5%, 99.08%, and 101.6%, respectively. The contents of total phenols and total saponins were 3. 04% -11. 9% and 0. 87% -3. 63%. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside fluctuated from 0.018% to 0. 572%, from 0.041% to 1.75% and from 0.035% to 1.32%. The established methods were reproducible, and they could be used for the quality control of Sargentodoxae Caulis. The present investigation suggested that total phenols, salidroside, and chlorogenic acid should be recorded in the quality standard of Sargentodoxae Caulis and their contents should not be less than 6.8% for total phenols, 0.040% for salidroside, and 0.21% for chlorogenic acid.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Magnoliopsida ; chemistry ; Phenol ; analysis ; Plant Stems ; chemistry ; Saponins ; analysis ; Triterpenes ; analysis