OBJECTIVETo explore a new method for early avascular necrosis of femoral head (AVNFH) therapy.

METHODSSixty-nine AVNFH New Zealand adult rabbits were randomly divided into three groups with equal number. In Group A, deproteinized bone (DPB) that absorbed with recombinant plasmid pcDNA3.1-hVEGF165 was implanted into the drilled tunnel of necrotic femoral head. In Group B, only DPB was implanted. In Group C, only tunnel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained at postoperative 1, 2, 4, 8, 16 weeks. The expression of VEGF165 and collagen I was detected by immunohistochemistry. Bone formation was detected generally by X-ray. Angiogenesis and the repair of the femoral head were observed histologically.

RESULTSThe expression of VEGF 165 could be detected 2 weeks after implantation in Group A, but it was not observed in other groups. The result of collagen I expression had a significantly difference 2, 4 and 8 weeks after operation in Group A from those in other groups (P < 0.01). X-ray results indicated that there was more bone formation in Group A than in other groups. The regenerated capillary vessels staining result of necrotic femoral head in Group A was significantly different from those in other groups at postoperative 2 and 4 weeks (P < 0.01).

CONCLUSIONSTransfection of hVEGF165 gene enhances local angiogenesis and DPB-VEGF compound improves the repair of necrotic femoral head. Deproteinized bone grafting with VEGF gene transfer provides a potential method for the treatment of osteonecrosis.

Animals ; Bone Transplantation ; Collagen Type I ; analysis ; Femur Head Necrosis ; pathology ; physiopathology ; therapy ; Genetic Therapy ; Immunohistochemistry ; Neovascularization, Physiologic ; Rabbits ; Transfection ; Vascular Endothelial Growth Factor A ; analysis ; genetics

OBJECTIVETo study the effect of peptide-conjugated near-infrared quantum dots (QDs) on growth, invasion and metastasis of human buccal squamous cell carcinoma cell line (BcaCD885 cell).

METHODS(1) BcaCD885 cells were labeled by cell-penetrating peptide-conjugated QDs with a maximum emission wavelength of 800 nm (QD800), then labeling efficiency was detected by flow cytometry, and laser-scanning confocal microscope was used to observe the distribution of QD800 within the cells. (2) Different concentrations of QD800 was applied to BcaCD885 cells, and the cell growth of control and three test groups were compared respectively. (3) BcaCD885 cells were labeled by QD800 (BcaCD885/QD800), then transwell chambers and wash way were used to detect the difference of invasion and metastasis ability between BcaCD885/QD800 and BcaCD885 cells.

RESULTS(1) The labeling rate of BcaCD885 cells after 6h was 94.07%, and QD800 distributes in the BcaCD885 cytoplasm. (2) Different concentrations of QD800 showed no negative effects on growth of BcaCD885 cells. (3) The ability of invasion, attachment and motion of BcaCD885 cells were not significantly different between test and control group (P > 0.05).

CONCLUSIONQDs showed no effects on growth, invasion and metastasis ability of BcaCD885 cells. Our results provide science foundation for QDs as a new fluorescence probes to real-time monitor cells and cells imaging in a living.

Animals ; Carcinoma, Squamous Cell ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Mice ; Mice, Nude ; Peptides ; Quantum Dots

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Objective To observe the effect of peptide-conjugated quantum dots with a maximal emission of 655 nm (QD655) on growth, proliferation, apoptosis and lymphatic metastasis of human tongue squamous cell carcinoma cell line Tca8113 and mouse uterine cervix carcinoma U14 in vivo.Methods Tca8113and U14 cells were labeled by QD655 (Tca8113-QD655, U14-QD655) .Tca8113-QD655 and U14-QD655 were inoculated subcutaneously into nude mice and Kunming mice.The tumor formation of Tca8113-QD655 and Tca8113 ,U14-QD655 and U14 was observed and compared in vivo.The proliferation and apoptosis of Tca8113-QD655 and Tca8113, U14-QD655, and U14 cells from tumors formed in vivo were analyzed by flow cytometry.U14-QD635 and U14 were inoculated into the buccal mucosa of Kunming mice to establish the cervical lymph node metastasis model of buccal cancer.The cervical lymph node metastatic ability of U14-QD655 and U14 was compared.Results The tumor weight and volume of Tca8113-QD655 and Tca8113, U14-QD655 and U14 in vivo were not significantly different (P > 0.05) , the cell proliferation index and apoptosis index of Tca8113-QD655 and Tca8113,U14-QD655 and U14 in vivo were not significantly different (P>0.05).The cervical lymph node metastasis rate of U14-QD655 and U14 buccal cancer were not significantly different (P > 0.05).Conclusions QD showed no effects on tumorigenicity and lymph node metastasis of Tca8113 and U14 cells.These results provide the scientific basis for noninvasive imaging and long-term tracing study.

Objective To examine the in vivo visual imaging of buccal carcinoma with the nearinfrared fluorescent quantum dots. Methods The U14 cells were labeled by endocytosis with QD800 (U14/QD800) which was linked with cell-penetrating peptide. Different number of U14/QD800 was injected under the buccal mucosa of nude mice and Kunming mice separately and imaged at different time to detect the in vivo sensitivity and dynamic imaging of U14/QD800. Results The minimum number of U14/QD800 cells which could be detected by in vivo imaging system was 1 × 104 in nude mice's cheek and 1 × 105 in Kunming mice's. The time for visual imaging of 1 × 104, 1 × 105 and 1 × 106 U14/QD800 cells in nude mice was 3, 7 and 16 d separately, and 3 and 10 d separately in Kunming mice. Conclusions Due to its strong tissue penetration, near-infrared fluorescent quantum dots have great prospects in cancer early diagnosis, visual observation and individual treatment.

OBJECTIVETo compare the therapeutic effects of aspiration via a directional soft tube and conservative treatment in patients with mild hemorrhage in the basal ganglion.

METHODSSeventy-five patients with mild cerebral hemorrhage (10~30 ml) were randomly divided into two groups for aspiration treatment with minimally invasive directional soft tube placement (minimally invasive group, n=36) and conservative treatment (medication group, n=39). The patients in the two groups had comparable mean GCS scores of 11-15 on admission. The clinical outcomes of the patients were compared between the two groups.

RESULTSIn the minimally invasive group, complete removal or absorption of the hematoma occurred within an average of 3.8 days, significantly shortened in comparison with the 24 days in the medication group. The short-term (1 month) follow-up of the patients showed good neurological recovery in 58% of the patients in the minimally invasive group, significantly greater than the rate of 29% in the medication group; 6 months after the treatment, good neurological recovery was achieved in 50% of the patients in the minimally invasive group, but only 16% in the medication. No death occurred in the minimally invasive group, and 2 patients died in the medication group. The cost of hospitalization averaged 5136.3 Yuan in the minimally invasive group and 11843.6 Yuan in the medication group.

CONCLUSIONCompared with conservative treatment, the minimally invasive treatment with soft tube placement can significantly shorten the hospital stay, promote neurological function recovery, lower the mortality rate, and reduce the cost of hospitalization.

Adult ; Aged ; Basal Ganglia Hemorrhage ; etiology ; surgery ; Catheters, Indwelling ; Female ; Humans ; Hypertension ; complications ; Male ; Middle Aged ; Suction ; economics ; methods ; Treatment Outcome

Objective To measure the avidity of serum autoantibodies to Aβs in patients with Alzheimer's disease (AD). Methods Enzyme-linked immtmoserbent assay (ELISA) combined with thiocyanate elution technique was employed to detect the avidity of serum autoantibodies to Aβs in patients with AD, middle-aged and healthy elderly adults (n=20). Results The avidity of serum autoantibodies to Aβs in patients with AD (avidity index, 1.6 [1.15 to 2.05]) was significantly lower as compared with that in healthy elderly subjects (avidity index, 2.45 [1.75 to 3.08]) (P=0.020) and no significant difference was showed in the avidity of autoantibodies to Aβs between the elderly and middle-aged healthy adults (P=0.221). An evident shift to the low section was observed in patients with AD in the avidity distribution histogram. The proportion of low affinity antibodies was significantly higher in patients with AD (13% [5% to 18%]) than that in healthy elderly subjects (5% [3% to 10%]) (P =0.000), and the proportion of high affinity antibodies was significantly lower in patients with AD (15% [5% to 24%]) than that in elderly adults (35% [26% to 44%]) (P=0.006). Conclusion Low avidity of autoantibodies to Aβs is confirmed in AD patients. Patients with AD show a significantly high proportion of low affinity antibodies than normal adults and the components of polyclonal antibodies change in patients with AD, probably resulting from incomplete immune tolerance of B cells.

OBJECTIVETo investigate the sensitivity of imatinib mesylate (IM) on Sup-B15 Ph⁺ acute lymphoblastic leukemia (ALL) cells knockdown of VE-cadherin (CD144), and to further explore its mechanism.

METHODSCD144 in Sup-B15 leukemia cells was stably knock downed via lentivirus-mediated RNA interference (named as Sup-B15/shVEC). The inhibitory effects of IM on Sup-B15/shVEC and Sup-B15 leukemia cells were measured by CCK-8 test, and the apoptosis of those cells was determined by AnnexinV/7-AAD dyeing using flow cytometry, the percentage of CD34⁺CD38⁻ leukemia cells also by flow cytometry. ALDH1 mRNA levels were detected by real-time RT-PCR, and protein levels of CD144, CD133, Bcr-abl and β-catenin by Western blot.

RESULTSIM treatment presented inhibitory effects on Sup-B15/shVEC and Sup-B15 leukemia cells at multiple concentrations of IM. The IC50 of IM on Sup-B15/shVEC and Sup-B15 leukemia cells were 25.1μmol/L and 18.7μmol/L, respectively (P<0.05). After 48h of 20 μmol/L IM treatment, the percentages of apoptosis cell in Sup-B15/shVEC cells and Sup-B15 cell were (13.52±2.06)% and (3.03±0.72) %, respectively (P<0.05). The percentage of CD34⁺CD38⁻ cells in Sup-B15 cells was significantly higher than in Sup-B15/shVEC cells [(2.39±0.28)% vs (0.96±0.07)%, P<0.05). As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/shVEC was remarkably downregulated, and the CD133 protein level was also downregulated in Sup-B15/shVEC cells. Both cytoplasmic and nucleic β-catenin protein levels (but not for Bcr-abl levels) decreased in Sup-B15/shVEC cells as compare to Sup-B15 cells.

CONCLUSIONKnockdown of CD144 sensitized Sup-B15 Ph+ ALL cells to IM. The possible mechanisms underlying this phenomenon might be via inhibiting β-catenin nucleic translocation and facilitating β-catenin degradation.

Antigens, CD ; genetics ; Benzamides ; pharmacology ; Cadherins ; genetics ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Endothelium, Vascular ; drug effects ; metabolism ; Gene Knockdown Techniques ; Humans ; Imatinib Mesylate ; Piperazines ; pharmacology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Pyrimidines ; pharmacology ; RNA Interference ; beta Catenin ; metabolism

OBJECTIVETo detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests.

METHODSA nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA.

RESULTSTwo positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR.

CONCLUSIONIt might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Humans ; Middle Aged ; RNA, Viral ; genetics ; metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SARS Virus ; genetics ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; diagnosis ; virology ; Young Adult