BACKGROUND:Panax notoginseng saponins promotes bone repair by improving vascular proliferation. Therefore, the scaffolds carrying panax notoginseng saponins were supposed to be used to improve bone repair at the bone defect region. However, the biocompatibility of scaffolds remains unclear.
OBJECTIVE:To evaluate the biocompatibility of panax notoginseng saponins-hydroxyapatite/chitosan scaffolds.
METHODS:A new bone repair scaffold has been generated by thoroughly mixtures of 0, 0.1, 1, 10 mg panax notoginseng saponins and chitosan/hydroxyapatite using in-situ composite technique and freeze-drying technique. Morphology and mechanical property of the scaffold were observed under a scanning electron microscope. (1) Cel proliferation test:rabbit bone marrow mesenchymal stem cel s of passage 3 were cultured in four kinds of drug loaded hydroxyapatite/chitosan composite material leaching liquor. Cel s normal y cultured were considered as controls. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide was used to measure absorbance value of cel s in each group. (2) Hemolysis test:Rabbit anticoagulated blood was added with four kinds of drug loaded hydroxyapatite/chitosan composite material leaching liquor. Absorbance values were measured using a microplate reader in each group. (3) Pyrogen test:The four kinds of drug loaded hydroxyapatite/chitosan composite material leaching liquor and saline were respectively injected into ear vein of rabbits, and the increase of rabbit body temperature was detected.
RESULTS AND CONCLUSION:Drug loaded hydroxyapatite/chitosan composite material contained three dimensional porous structure of 110μm in diameter. Drug loading process of panax notoginseng saponins did not significantly affect the porosity, pore size and density of the composite material, but decreased its breaking strength and elastic modulus. The larger amount of drug loading showed a more obvious effect. Simple hydroxyapatite/chitosan composite material had good cel ular compatibility. The composite material after drug loading obviously suppressed cel proliferation, and the larger amount of drug loading showed a more obvious effect. The composite material had good blood compatibility before and after drug loading. The composite material had good pyrogen effects before and after drug loading, but accorded with acceptable quality level of pyrogen test.

In this study, we investigated the effect of aspirin on tumor biological effects mediated by hepatocyte growth factor/cellular-mesenchymal-epithelial transition factor (HGF/c-Met) axis, and preliminarily explored the molecular mechanism of inhibiting tumor metastasis by aspirin. The binding of aspirin to c-Met was predicted by molecular docking; cellular thermal shift assay (CETSA) was used to verify the binding of aspirin to c-Met at the cellular level. The inhibitory effect of aspirin on c-Met kinase was detected by kinase activity; Western blot, cell scattering test, cell branching morphogenesis and Transwell test were used to evaluate the cell signal transduction, morphological changes and migration and invasion ability. The results showed that aspirin could effectively inhibit the kinase activity of c-Met with a half inhibitory concentration of 0.95 mmol·L^-1. The results of docking showed that aspirin could bind to the ATP pocket of c-Met protein, and the main binding sites were Tyr1230, Tyr1159 and Met1229. The CETSA test also showed that aspirin could form binding complex with c-Met protein. Western blot results showed that aspirin could inhibit the up-regulation of phosphorylated Met stimulated by HGF in a concentration-dependent manner. The results of cell scattering test showed that aspirin could block HGF/c-Met promoted cell scattering in a concentration dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. Similarly, the results of MDCK cell branching morphogenesis experiment showed that aspirin could inhibit HGF/c-Met mediated invasive growth in a concentration dependent manner. The results of Transwell test showed that aspirin could block HGF/c-Met mediated cell migration and invasion in a concentration-dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. The above results indicate that aspirin can bind to c-Met, thereby blocking the biological effects mediated by HGF/c-Met, and inhibiting tumor metastasis. This study revealed the new biological function of aspirin, and provided a new theoretical basis for a comprehensive understanding of the anti-metastatic effect of aspirin.

Objective To investigate the expression and genetic alterations of KLF6 in hepatocellular carcinoma (HCC) and explore their functional mechanisms in the oncogenesis and development of HCC. Methods Real-time quantitative-PCR, direct sequencing and LOH approaches were used to detect KLF6 genetic abnormalities in HCC. The experiment had 2 groups, an experimental group and a control group. In the experimental group, the transfected plasmid pcDNA3.0 was recombined with KLF6 and tranfected into HCC HepG2 cells. MTT, flow cytometry and Western blotting were used to observe the effect of anti-oncogene wild type KLF6 on HepG2 cells by transgenic method for 48 h.Results Expression levels of KLF6 were significantly downregulated in HCCs(P＜0. 01), as detected by qRT-PCR. LOH occurred in 11 (52%) of the 21 tumors, and all the samples with LOH showed KLF6 down-regulation. The mutational frequency was 29%, and sequence changes located in activation domain of KLF6. Meanwhile, MTT assay showed a significant antiproliferative effect of the transfected wtKLF6 on HepG2 cells(42.7%, P＜0.05). Fluorescence-activated cell sorting analysis revealed that KLF6 induced apoptosis. Conclusion The deregulation of KLF6 together with genetic abnormalities of allelic imbalance and mutations may play an important role in HCC pathogenesis.

Circular dichroism (CD) is an useful technique for monitoring DNA conformation changes resulting from changes in environmental conditions, such as temperature, ionic strength, and pH, and also for the study of the interaction between DNA and ligands (including small molecules and proteins). CD spectroscopy of DNA arises from the asymmetric backbone sugars and by the helical structures often adopted by nucleic acids. By the interpretation of induced circular dichroism (ICD) of ligand signals resulting from the coupling of electric transition moments of the ligand, DNA bases within the asymmetric DNA environment, ligand-DNA interactions, as well as the DNA-binding mode can be assessed. A number of important conclusions have been reported that related to the observed ICD signals resulting from the interactions between intercalators and groove binders with DNA. If short oligonucleotide sequences are used in the study, sequences-specific of binding also can be deduced. CD determination requires smaller amounts of sample, and not limited by the molecular weight or size and can be performed rapidly; though CD is of low resolution, but it's a complement to NMR and X-ray diffraction methods. This review will introduce the characters of the CD spectra of DNA, and its application to the studies of DNA with small molecules; some progress of the studies in our laboratory will also be discussed. CD is expected to be used as a screening method in seeking more DNA-targeted drugs, such as, antineoplastic, antimicrobial and antiviral drugs.
Animals ; Antineoplastic Agents ; chemistry ; Base Sequence ; Circular Dichroism ; methods ; DNA ; chemistry ; metabolism ; Humans ; Intercalating Agents ; chemistry ; Ligands ; Protein Binding ; Small Molecule Libraries ; pharmacology

OBJECTIVETo identify the mutation of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis (ALS) family with unique phenotype.

METHODSFive exons of SOD1 gene were amplified by PCR. The differences of these products were analyzed by PCR-single strand conformation polymorphism and visualized by silver staining.

RESULTSAbnormal bands were found in exons 2 and 5 of SOD1 gene in several familial members. DNA sequence analysis verified that a base pair insertion occurred in the codon area of exon 2 and in the intron area of exon 5. And the insertion mutation of exon 2 led to a frameshift mutation and premature stop. It is a new type of SOD1 mutation which may be associated with familial amyotrophic lateral sclerosis.

CONCLUSIONInsertion mutation of exon 2 may be responsible for the disease of an ALS family in Chongqing.

Adult ; Amyotrophic Lateral Sclerosis ; genetics ; Humans ; Mutation ; Polymorphism, Single-Stranded Conformational ; Superoxide Dismutase ; genetics ; Superoxide Dismutase-1

OBJECTIVETo explore the clinical efficacy of the percutaneous balloon kyphoplasty for osteoporotic vertebral compression fractures with osteonecrosis.

METHODSThe clinical data of 31 patients with osteoporotic vertebral compression fractures associated with osteonecrosis from January 2005 to January 2008 were analyzed retrospectively. There were 13 male and 18 female in this study. The mean age of the patients was 71 years (range from 57 to 84 years). The back pain lasted for 4.2 months (from 1 month to 10 years). Radiography, MRI and CT examination were performed. The patients were treated by percutaneous balloon kyphoplasty and the vertebral body tissue was extracted to perform common pathological examination. The anterior vertebral height was measured on a standing lateral radiograph before operation, after operation (one day after operation) and at the final follow-up. A Visual Analog Scale (VAS) and the Oswestry Disability Index (ODI) were chosen to evaluate pain status and functional activity.

RESULTSThe mean follow-up was for 27 months (range, 18 to 48 months). The anterior vertebral body height of fracture vertebra was restored from (34.7 +/- 3.1)% preoperatively to (71.4 +/- 2.3)% postoperatively, and to (70.2 +/- 2.5)% at the final follow-up. There was a significant improvement between preoperative and postoperative values (P < 0.05) and no difference between postoperatively and at the final follow-up (P > 0.05). The VAS was 8.7 +/- 0.4 preoperatively, 2.3 +/- 0.7 postoperatively, and 1.9 +/- 0.2 at the final follow-up; and the ODI was 89.1 +/- 2.7 preoperatively, 31.7 +/- 3.1 postoperatively, and 29.1 +/- 2.7 at the final follow-up. There was statistically significant increment in the VAS and ODI postoperatively compared with preoperatively (P < 0.05), while there was no statistically significant differences between postoperatively and at the final follow-up (P > 0.05). There was a significant increment between preoperative and final follow-up values (P < 0.05). Asymptomatic cement leakage occurred in two cases. New vertebral fracture occurred in one case.

CONCLUSIONBalloon kyphoplasty is a safe and effective procedure for osteoporotic vertebral compression fractures with osteonecrosis.

Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Fractures, Compression ; complications ; etiology ; Humans ; Kyphoplasty ; methods ; Male ; Middle Aged ; Osteonecrosis ; etiology ; surgery ; Osteoporosis ; complications ; Retrospective Studies ; Spinal Fractures ; complications ; etiology ; Vertebroplasty

OBJECTIVE To investigate the surgical methods for the lesions involved the common carotid artery.METHODS The clinical data of 11 cases with lesions involved the common carotid artery who underwent operations were retrospectively studied.The lesions were 1 case with recurrence tumor after 3/4 partial laryngectomy,1 case with bleeding of the carotid aneurysm caused by tuberculosis,1 case with iatrogenic carotid aneurysm,3 cases with carotid body tumor,1 case with thyroid gland cancer,2 cases with neck tumor,1 case with injury of the carotid artery and 1 case with gas gangrene.RESULTS Common carotid artery was reconstructed in 2 cases after removal of the tumors.The tumors were resected using the carotid shunt in 2 cases.Common carotid artery was sutured in 1 case with neck injury.The common carotid artery was repaired in 1 case with iatrogenic carotid aneurysm after removal of the tumor.The carotid artery was dissected out from the thyroid gland cancer in 1 case.The common carotid artery was reserved in 2 cases after resection of the neck tumors. Neck drainage was performed in the case with gas gangrene.CONCLUSION The surgical methods for lesions involved the carotid artery after removal of the tumors include the reconstruction of the carotid artery, resection and suture the carotid artery,and free of the carotid artery from the tumors.

Objective To study the impacts of HIV/HCV co-infection to NK cells by investigating the changes of frequencies and functions of NK cells in HIV/HCV co-infected patients. Methods Frequencies and counts of NK cells were measured in patients with HIV mono-infection, HCV monoinfection, HIV/HCV co-infection and health control (HC) group by flow cytometer ( FCM ). After stimulated with PMA and K562 cells, PBMCs were examined the proportion of IFN-γ+ NK cells by FCM. Proportion of killed K562 cells were detected to analyze the killing functions of NK cells. Results The frequencies of NK cells, the percentages of IFN-γ+ NK cells as well as the functions of NK cells killing the K562 cells in HIV/ HCV co-infection, HIV mono-infection and HCV mono-infection groups were all lower than those of HC group significantly, the absolute counts and killing functions of NK cells in co-infection group were significantly lower than those of HIV or HCV mono-infection group. Conclusions The counts and functions of NK cells were affected more in HIV/HCV co-infections than those in HIV or HCV mono-infection.

OBJECTIVETo investigate the application of the island flap based on the postfemur neurocutaneous nutrient vessel.

METHODSThe flap was designed and applied to repair the defects in the gluteal, popliteal fossa or the bilateral postfemur areas. A total of 11 cases (12 defects) were treated with this method. The size of the defects ranged from 4.0 cm x 7.8 cm to 8.3 cm x 16.6 cm.

RESULTSOf the 12 defects, 9 achieved complete success. Epidermal necrosis occurred in the distal part of the flap in 3 defects owing to venous stasis, which were cured with skin grafting. Postoperative follow-up for 8-19 months showed that the appearance, texture, and function of the flap were satisfactory.

CONCLUSIONSThe advantages of the flap lie in the reliable blood supply, constant anatomy, and without sacrificing a major artery. The key points for the flap survival are utilizing the "Superficial vein-nutrient vessel of the cutaneous nerve system" and retaining a comet tail-shaped soft-tissue pedicle in the flap creation.

Arteries ; Follow-Up Studies ; Humans ; Necrosis ; etiology ; surgery ; Skin ; injuries ; pathology ; Skin Transplantation ; Surgical Flaps ; blood supply ; pathology ; transplantation ; Thigh ; Wound Healing

OBJECTIVETo evaluate the efficacy and safety of a China made adefovir dipivoxil (ADV) treatment for hepatitis B e antigen-positive patients with chronic hepatitis B.

METHODSTwo hundred and thirty patients with chronic hepatitis B who were positive for hepatitis B e antigen (HBeAg) were randomly put into groups A or B, and 58 patients with lamivudine-resistant chronic hepatitis B were randomly put into groups C or D. During the first 12 weeks of the trial, 112 patients in group A and 115 patients in group B received 10 mg of ADV and a placebo once a day; 28 patients in group C received 100 mg of lamivudine (LMV) and 10 mg of ADV; 29 patients in group D received 100 mg of LMV and a placebo once a day. In the second trial period, all patients received ADV for 36 weeks. The primary checking criterion was the serum HBV DNA change during the treatment. The secondary ones were alanine aminotransferase (ALT) normalization, HBeAg loss, and HBeAg seroconversion.

RESULTSAt week 12, the median serum hepatitis B virus (HBV) DNA level of group A (ADV-ADV) was reduced 2.8 log10 copies/ml, significantly greater than that of group B (placebo-ADV) of 0.3 log10 copies/ml reduction (P = 0.000). At week 48, the median serum HBV DNA level of group A and group B were reduced 3.6 and 3.4 log10 copies/ml respectively. At week 12, the median serum HBV DNA level of group C (LMV+ADV) was reduced 3.0 log10 copies/ml, significantly greater than that of the group D (LMV+placebo) of 0.16 log10 copies/ml reduction (P = 0.000). At week 48, the median serum HBV DNA level of group C and group D were reduced 3.6 and 3.8 log10 copies/ml respectively. Only 5.56% (16/288) patients had adverse events that were mild to moderate. There was no significant difference in the change of serum creatinine compared with their baseline levels.

CONCLUSIONIn our HBeAg positive lamivudine-resistant chronic hepatitis B patients, 48 weeks of ADV treatment was safe and resulted in significant virological and biochemical improvements.

Adenine ; analogs & derivatives ; therapeutic use ; Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Double-Blind Method ; Drug Resistance, Viral ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; immunology ; virology ; Humans ; Middle Aged ; Mutation ; Organophosphonates ; therapeutic use ; Young Adult