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 discuss the radical treatment of cervical giant cystic lymphangioma in Children and cosmetic result.

METHODSTwenty-five children with cervical giant cystic lymphangioma were retrospectively analyzed. The diameter of all the tumors was more than 10 cm. 24 cases underwent resection. The complication, therapeutic effect and cosmetic result were recorded.

RESULTSThe tumors were all removed radically in all the cases. The patients were followed up for 1-5 years with no recurrence. Cosmetic result was satisfactory in 22 cases. Secondary operation was performed in 2 cases with satisfactory result. Complications included 5 cases of lymph leakage, 2 cases of poor wound healing, 1 case of infection and 2 cases of tongue edema.

CONCLUSIONSThe cervical giant cystic lymphangioma in children can be resected radically with satisfactory result.

Child, Preschool ; Female ; Head and Neck Neoplasms ; surgery ; Humans ; Infant ; Lymphangioma, Cystic ; surgery ; Male ; Neck ; Retrospective Studies ; Treatment Outcome

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

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

Aim To investigate the protective effects of supercritical CO2 fluid extract(SFE)of Notoginseng a-gainst glutamate-induced PC12 cells damage and the underlying mechanism. Methods PC12 cells were dealt with glutamate to establish cell models. MTT as-say,LDH method,Hoschst 33342 staining,Fluo-3 /AM fluorescence staining and Western blot were used to observe the changes of cell viability,intracellular Ca2 + concentration and the expression of protein that interacted with C kinase l(PICK1)and glutamate re-ceptors 2 (GluR2),respectively. Results Glutamate was cytotoxic to PC12 cells with an inhibitory concen-tration 50(IC 50 )of 25 mmol·L - 1 . Pretreatment with SFE(25,50,100 mg·L-1)and FSC231(100 μmol ·L-1 )and SFE(100 mg·L-1 )+FSC231(100μmol ·L-1 )remarkablely improved cell viability,reduced LDH leakage,decreased apoptosis rate,debased intra-cellular calcium concentration,decreased the expres-sion of PICK1 ,and increased the expression of GluR2 . Conclusions SFE of Notoginseng shows protective effects against glutamate-induced PC12 cell damage, and its mechanism may be related to the inhibition of PICK1 and the increase of GluR2 protein expression.

This study was purposed to construct lentivirus vector containing human homeobox gene HOXB4 and explore changes of human umbilical cord mesenchymal stem cells (HUCMSC) after infected with HOXB4 mediated by lentivirus. PCR amplification was performed to obtain HOXB4, which was cloned in lenti-shuttle vector. Four-plasmid lentivirus packaging system was used to transfect HEK293T cells. After 48 h, lentivirus Lenti-HOXB4 was harvested and lentivirus titer was determined. Lenti-HOXB4 was used to infect HUCMSC. The infected cells were observed under inverted fluorescence microscope to determine the optimal multiplicity of infection (MOI). Meanwhile, RT-PCR, immune fluorescence staining, CCK-8 and flow cytometry (FCM) were used to determine the expression of HOXB4 and its effect on cell growth. The results indicated that lenti-HOXB4 was successfully obtained by co-transfecting the 293T cells with four plasmids. The determined virus titer was 3×10(8) TU/ml; when MOI was 20. Lenti-HOXB4 had a high transfection rate in HUCMSC, over 80%. In HUCMSC infected with lenti-HOXB4, the expression of target gene could be detected both at mRNA and protein levels. It could promote the proliferation of HUCMSC. FCM results indicated HOXB4 gene did not significantly influence the surface marker of HUCMSC. It is concluded that HOXB4 gene can promote the high proliferation of HUCMSC and does not significantly influence the expression of the surface marker of HUCMSC.
Cell Proliferation ; Cells, Cultured ; Flow Cytometry ; Genes, Homeobox ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; cytology ; Plasmids ; Transcription Factors ; genetics ; Umbilical Cord ; cytology

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

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

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