OBJECTIVETo introduce the postero- lateral surgical approach to the posterior malleolar fracture and report its clinical outcomes in 32 cases.

METHODSThis study consisted of 32 cases, 22 males and 10 females with the mean age of 48 years (range, 21-63 years), suffering from posterior malleolar fracture. All cases were treated with the posterolateral surgical approach to the ankle. The average follow-up period was 28 months (range, 24-35 months). The clinical outcomes of these cases were evaluated on the basis of the Olerud-Molander Ankle (OMA) score and plain radiographs.

RESULTSAll cases showed radiological evidence of bony union at follow-up. The average OMA score was 82 points; 21 cases had excellent scores (90-100 points), 9 good (61-90 points), and 2 fair (31-60 points). The excellent-to-good rate was 93.8%. Although most cases did not show any wound dehiscence or necrosis, one patient had a superficial infection which healed after using antibiotic dressing and one had sural cutaneous nerve injury that underwent spontaneous remission without any treatment after three months. In addition, one presented with mild symptoms of peroneal tendonitis that disappeared after plate removal.

CONCLUSIONThe posterolateral approach offers an effective technique for fracture reduction and fixation of large posterior malleolar fragments.

Ankle Fractures ; Ankle Joint ; surgery ; Bone Plates ; Fracture Fixation, Internal ; Humans

OBJECTIVETo isolate and identify the cancer stem cells from primary human ovarian cancer tissues.

METHODSFresh tumor tissues from five cases of pathologically diagnosed ovarian cancers were taken, minced and then digested with collagenase and hyaluronidase to obtain single cell suspension. The erythrocytes were removed with ACK Lysis buffer. The suspensions were sorted by magnetic activated cell sorting (MACS) using CD133-binding microbeads. Then the sorted CD133(+) cells were verified by flow cytometry. The cells were cultured in serum-free medium supplemented with EGF, bFGF, insulin and BSA, and grew into spheroids. Immunofluorescence, differentiation and tumor formation tests of the cells were performed to characterize the properties of cancer stem cells.

RESULTSThe ovarian cancer stem cells were successfully isolated from primary human ovarian tumors, which formed typical spheroids in serum-free medium and had stronger ability of tumorigenesis. The results of related experiments verified that CD133 positive cells owned the properties of cancer stem cells.

CONCLUSIONSThe ovarian cancer stem cells presenting the characteristics of stemness in vitro and in vivo, have been successfully isolated from primary human ovarian tumor tissues by MACS. The isolated ovarian cancer stem cells could be used in future researches of their biological functions.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Cell Differentiation ; Cell Separation ; methods ; Female ; Flow Cytometry ; methods ; Glycoproteins ; metabolism ; Humans ; Immunomagnetic Separation ; methods ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Peptides ; metabolism

Objective To measure albumin in urine by HPLC and conduct primary clinical application Methods Solvent gradient and appropriate wave length was optimized and performance of the HPLC method was evaluated.Urine albumin of 46 patients with diabetes was measured.Results In standard and urine,retention time of Alb was 13.1 min.The linear measuring range extends to 1 820 mg/L.The lower limit of measurment for Alb was 4.2 mg/L.The intra-assay CV and the inter-assay CV were 3.36%,4.12% and 1.93%,1.97% at 24.5 mg/L and 546.9 mg/L of Alb respectively.Analytical recovery rate were 96.3%,98.2% and 97.5%.Microalbuminuria rate was 54.3% by HPLC,26.1% by immunoassay in 46 patients with diabetes.Conclusions Measurement of Alb in urine by HPLC is feasible as routine method until quantifying urinary total Alb conveniently.HPLC is the same to suit research for diabetic nephropathy and so on.

BACKGROUNDMany studies have shown the superior efficacy of budesonide (BUD)/formoterol (FORM) maintenance and reliever therapy, but still lack evidence of its efficacy in Chinese asthma patients in a relative large patient-group. We finished this research to compare BUD/FORM maintenance and reliever therapy and high-dose salmeterol (SALM)/fluticasone (FP) maintenance plus an as-needed short-acting β(2)-agonist in Chinese patients with persistent uncontrolled asthma. This was a post hoc analysis based on a 6-month, multicenter, randomized, double-blind study (NCT00242775).

METHODSA total of 222 eligible asthma patients from nine centers in China were randomized to either BUD/FORM+as-needed BUD/FORM (160/4.5 µg/inhalation) (640/18 µg/d; n = 111), or SALM/FP+as-needed terbutaline (0.4 mg/inhalation) (100/1000 µg/d; n = 111). The primary endpoint was time to first severe exacerbation while secondary endpoints included various measures of pulmonary function, symptom control and quality-of-life.

RESULTSTime to first severe exacerbation over six months was lower with the BUD/FORM than with the SALM/FP treatment (risk ratio = 0.52, 95%CI 0.22 - 1.22), but the difference did not achieve statistical significance (P = 0.13). The cumulative number of severe exacerbations in the BUD/FORM group was lower than in the SALM/FP group (7.2% vs. 13.5%; risk ratio = 0.45, P = 0.028). BUD/FORM produced significantly better improvements in reliever use, cumulative mild exacerbations, symptom-free days (%), and morning/evening peak expiratory flow (PEF) than SALM/FP (P < 0.05 in all cases). The two groups achieved similar improvements in their time to first mild exacerbation, forced expiratory volume in one second (FEV(1)), asthma control questionnaire and asthma symptom scores, and percentage of nights with awakening(s). Both treatments were well tolerated.

CONCLUSIONSIn Chinese patients with persistent asthma, BUD/FORM decreased severe and mild exacerbations, decreased reliever use, increased symptom-free days, and improved morning/evening PEF compared with SALM/FP. There were no significant differences in time to first severe exacerbation or other assessments regarding daily asthma control between BUD/FORM and SALM/FP. BUD/FORM was more effective in this Chinese sub-group than in the total cohort involved in the original study.

Adolescent ; Adult ; Aged ; Asthma ; complications ; drug therapy ; physiopathology ; Budesonide ; administration & dosage ; adverse effects ; Double-Blind Method ; Ethanolamines ; administration & dosage ; adverse effects ; Female ; Forced Expiratory Volume ; Formoterol Fumarate ; Humans ; Male ; Middle Aged

BACKGROUNDNuclear factor kappaB (NF-kappaB) overactivation, requiring phosphorylation and degradation of its inhibitor IkappaBalpha, is the basis for chronicity of airway inflammation in asthma. Based on our previous plasmid pShuttle-IkappaBalpha, carrying an IkappaBalpha gene from human placenta, we optimized a novel IkappaBalpha mutant (IkappaBalphaM) gene, constructed and characterized its replication-deficient recombinant adenovirus (AdIkappaBalphaM), and tested whether AdIkappaBalphaM-mediated overexpression of IkappaBalphaM could inhibit the NF-kappaB activation in endothelial cells.

METHODSIkappaBalphaM gene (203 - 1003 bp) encoding 267 amino acids, acquired by site-directed deleting N-terminal phosphorylation sites of serine 32/36, was subcloned into the pShuttle and pGEM-T vectors for further polymerase chain reaction (PCR), restriction digestion, deoxyribonucleic acid (DNA) sequencing and homology analyses. Subsequent to inserting the expression unit of pShuttle-IkappaBalphaM, containing cytomegalovirus (CMV) promoter, IkappaBalphaM complementary DNA (cDNA) and polyadenylic acid (PolyA) signals, into the type 5 adenovirus (Ad5) vector, the resultant AdIkappaBalphaM was packaged in human embryonic kidney (HEK) 293 cells by cotransfection with lipofectamine. Western blot analysis and electrophoretic mobility shift assay were utilized to detect the AdIkappaBalphaM-mediated overexpression of IkappaBalphaM in HEK293 cells and its suppressive effect on phorbol 12-myristate 13-acetate (PMA)-induced NF-kappaB activation in human umbilical vein endothelial (ECV304) cells, respectively.

RESULTSThe relevant nucleotides and deduced amino acids of 801 bp IkappaBalphaM gene were consistent with those of IkappaBalpha gene (GenBank accession number: M69043). The titer of the prepared AdIkappaBalphaM was 4.0 x 10 (12) plaque-forming units (pfu)/L. Moreover, the IkappaBalphaM gene was overexpressed in HEK293 cells, and potently inhibited the PMA-induced NF-kappaB activation in ECV304 cells dose-dependently.

CONCLUSIONSAdIkappaBalphaM is a novel vector for both efficient transfer and specific overexpression of IkappaBalphaM gene, as well as potent inhibition of NF-kappaB activity, providing a promising strategy for gene therapy of asthma.

Adenoviridae ; genetics ; Cell Line ; Endothelial Cells ; metabolism ; Genetic Therapy ; Humans ; I-kappa B Proteins ; genetics ; Mutation ; NF-KappaB Inhibitor alpha ; NF-kappa B ; antagonists & inhibitors ; Tetradecanoylphorbol Acetate ; pharmacology

OBJECTIVETo construct an adenovirus vector containing the double-mutant hypoxia-inducible factor-1alpha (HIF-1alpha) gene for exploring the therapeutic angiogenesis for coronary heart disease.

METHODSHuman double-mutant HIF-1alpha cDNA was obtained from PCR of pShuttle-2-HIF-1alpha containing the mutant HIF-1alpha gene (564). The recombinant adenoviral plasmid containing mutant HIF-1alpha cDNA was constructed by ligation of recombinant pShuttle2 containing double-mutant HIF-1alpha cDNA and Adeno-X viral DNA, followed by its identification and transfection into adenoviral packaging cells HEK293 via lipofectamine 2000.

RESULT AND CONCLUSIONThe recombinant pAdeno-HIF-1alpha was correctly constructed and verified by restriction endonuclease and DNA sequencing analyseis. This recombinant adenovirus containing the double-mutant HIF-1alpha gene may facilitate further investigation of mutant HIF-1alphagene therapy for coronary heart disease.

Adenoviridae ; genetics ; Cell Line, Tumor ; Coronary Disease ; therapy ; Genetic Therapy ; Genetic Vectors ; biosynthesis ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Plasmids

Ob jective It has been reported by some prospective studies that C-reactive protein (CRP ) is associated with cancer risk .However, the correlation between CRP and digestive system cancers has not been evaluated in Chinese females .We conducted a large population-based cohort study to investigate whether elevated level of CRP in serum is associated with an increased risk of digestive system cancers in Chinese women.Metho ds From the Chinese Kailuan Female Cohort , 19,437 women were enrolled in this study in July 2006, and all of the subjects were followed up through 2014.At the baseline investigation , the serum levels of high-sensitivity CRP ( hsCRP ) were tested for all subjects , and demographic information and risk factor data were collected .Multivariable Cox proportional hazards regression model was used to calculate the hazard ratios ( HR ) and 95%confidence intervals ( 95%CI ) for the baseline levels of hsCRP after adjusting for age, marital status, smoking, drinking, body mass index ( BMI), diabetes and physical activity, and risk of digestive system tumors (including colorectal cancer, stomach cancer, pancreas cancer, liver and gallbladder cancer, and other cancers).Results By Dec 31, 2014, a total of 100 incident cancer cases were observed , including 47 colorectal cancers , 17 stomach cancers , and altogether 29 pancreas , liver and gallbladder cancers .All the subjects investigated were divided into three groups according to the level of hsCRP (<1 mg/L, 1-3 mg/L and >3 mg/L) .The 8-year cumulative incidence of digestive system cancers were 405/100 000, 520/100 000 and 787/100 000 in these 3 groups, respectively (Log rank test χ2=8.37, P=0.015 ) .Compared to those with lower hsCRP levels (<1 mg/L ) , the women with higher hsCRP (>3 mg/L) had a significantly increased risk of pancreas , liver and gallbladder cancers ( HR =2.70, 95%CI =1.06-6.91;Ptrend=0.036).Conclusions Elevated levels of hsCRP at baseline may be associated with increased risk of certain digestive system cancers .

To investigate the crucial role of particle size in the biological effects of nanoparticles, a series of mesoporous silica nanoparticles (MSNs) were prepared with particle size gradients (50, 100, 150, 200 nm) with the traditional Stober method and adjusting the type and ratio of the silica source. The correlation between toxicity and size-caused biological effects were then further examined both in vitro and in vivo. The results indicated that the prepared MSNs had a uniform size, good dispersal, and ordered mesoporous structure. Hemolytic toxicity was found to be independent of particle size. At the cellular level, MSNs with smaller particle sizes were more readily internalized by cells, which initiated to more intense oxidative stress, therefor inducing higher cytotoxicity, and apoptosis rate. In vivo studies demonstrated that MSNs primarily accumulated in the liver and kidneys of mice. Pharmacokinetic analysis revealed that larger MSNs were eliminated more efficiently by the urinary system than smaller MSNs. The mice's body weight monitoring, blood tests, and pathological sections of major organs indicated good biocompatibility for MSNs of different sizes. Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Zhejiang Chinese Medical University. Overall, this study prepared MSNs with a particle size gradient to investigate the correlation between toxicity and particle size using macrophages and endothelial cells. The study also examined the biosafety of MSNs with different particle sizes in vivo and in vitro, which could help to improve the safety design strategy of MSNs for drug delivery systems.

Ob jective It has been reported by some prospective studies that C-reactive protein (CRP ) is associated with cancer risk .However, the correlation between CRP and digestive system cancers has not been evaluated in Chinese females .We conducted a large population-based cohort study to investigate whether elevated level of CRP in serum is associated with an increased risk of digestive system cancers in Chinese women.Metho ds From the Chinese Kailuan Female Cohort , 19,437 women were enrolled in this study in July 2006, and all of the subjects were followed up through 2014.At the baseline investigation , the serum levels of high-sensitivity CRP ( hsCRP ) were tested for all subjects , and demographic information and risk factor data were collected .Multivariable Cox proportional hazards regression model was used to calculate the hazard ratios ( HR ) and 95%confidence intervals ( 95%CI ) for the baseline levels of hsCRP after adjusting for age, marital status, smoking, drinking, body mass index ( BMI), diabetes and physical activity, and risk of digestive system tumors (including colorectal cancer, stomach cancer, pancreas cancer, liver and gallbladder cancer, and other cancers).Results By Dec 31, 2014, a total of 100 incident cancer cases were observed , including 47 colorectal cancers , 17 stomach cancers , and altogether 29 pancreas , liver and gallbladder cancers .All the subjects investigated were divided into three groups according to the level of hsCRP (<1 mg/L, 1-3 mg/L and >3 mg/L) .The 8-year cumulative incidence of digestive system cancers were 405/100 000, 520/100 000 and 787/100 000 in these 3 groups, respectively (Log rank test χ2=8.37, P=0.015 ) .Compared to those with lower hsCRP levels (<1 mg/L ) , the women with higher hsCRP (>3 mg/L) had a significantly increased risk of pancreas , liver and gallbladder cancers ( HR =2.70, 95%CI =1.06-6.91;Ptrend=0.036).Conclusions Elevated levels of hsCRP at baseline may be associated with increased risk of certain digestive system cancers .

Adolescent ; Adult ; Aged ; Blood Banks ; legislation & jurisprudence ; Blood Donors ; legislation & jurisprudence ; Child ; Female ; Hepatitis C ; epidemiology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult