BACKGROUND:Immortalized cervical epithelial cels H8 can become cancerous under the induction of carcinogenic agent, and may cause cervical cancer when there is a cofactor interaction. However, there is stil a lack of effective intervention for female patients with precancerous lesions, and this treatment is blank in the clinic. OBJECTIVE: To explore the effects and mechanism of astragalus injection on apoptosis of human immortalizedcervical epithelial cels H8. METHODS: This study contained two groups: astragalus drug group and the blank control group. (1) Enzyme linked immunosorbent assay (ELISA) was used to detect DNA fragments of apoptotic H8 after astragalus injection. (2) Enzyme-labeling instrument was used to analyze the changes in caspase-3 and caspase-9 activities a fter astragalus injection. (3) Western blot assay was used to detect the protein expression changes of caspase-3, caspase-9 and PARP in H8 cels after astragalus injection. RESULTS AND CONCLUSION: (1) ELISA results showed that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, DNA fragments were gradualy increased with time prolonged in a time-dependent effect (P < 0.05). (2) Enzyme-labeling instrument demonstrated that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, caspase-3 and caspase-9 activities increased in a time-dependent manner (P < 0.05). (3) At 0, 6, 12 and 24 hours after 20 g/L astragalus injection, the expression of cleaved caspase-3 and cleaved caspase-9 were gradualy increased in H8 cels (P < 0.05). Cleaved PARP protein expression was gradualy decreased (P < 0.05). These findings indicate that astragalus injection could obviously induce H8 apoptosis, which may be associated with the upregulated protein expression of caspase-3 and caspase-9.

OBJECTIVETo investigate the severity of early myocardial injury in rats with 30% full thickness burn at plateau and the protective effects of Rhadiola Astragalus Codonopsis Compound (RACC) on the rat myocardial injury.

METHODSOne hundred and four Wistar rats with 30% full thickness burn were randomly divided into RACC application (R, n = 48) and scalding group 1 (S, n = 48), and another 8 healthy Wistar rats as control group 2 (C, n = 8). Four ml of RACC was garaged into the rat stomach in R and 4 ml isotonic saline in S groups respectively, but no treatment in C group. Blood samples from the aorta were harvested in 3, 6, 12, 24, 48 and 72 postburn hours (PBH) for blood gas analysis and for the determination of the changes in myocardial enzymes. Rat heart was harvested for pathomorphological examination.

RESULTSThe rat myocardial tissue injury in R and S groups was obvious at 3 PBH and ameliorated gradually thereafter, up to the degree in C group at 72 PBH. The serum levels of myocardial enzymes in R and S groups were significantly higher than those in C group (P < 0.01). Whereas the enzymes in R group were much lower than those in S group (P < 0.01). It was indicated by blood gas analysis that the pH in R and S groups was lower than that in C group (P < 0.05), while that in R group at 12 - 24 PBH was higher than that in S group (P < 0.05). In addition, the base excess in R and S groups was lower than that in C group (P < 0.01), while that in R group at 6 PBH was higher than that in S group (P < 0.05 approximately 0.01). The PaCO2 in R and S groups was evidently lower than that in C group (P < 0.05 approximately 0.01), while that in R group at 48 PBH was no different to that in C group (35.70 +/- 4.23 mmHg vs 37.50 +/- 6.53 mmHg, P > 0.05). The PaO2 in R and S groups at 3 approximately 24 PBH was higher than that in C group and decreased gradually (P > 0.05). There was no difference in SaO2 among 3 groups (P > 0.05).

CONCLUSIONRACC exhibited beneficial to the protection of rat heart from myocardial injury at plateau induced by severe burn.

Animals ; Astragalus Plant ; Blood Gas Analysis ; Burns ; drug therapy ; Codonopsis ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart ; drug effects ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar

OBJECTIVETo evaluate the role of flow cytometry in semen assessment.

METHODSSemen samples from 104 infertile male patients (as the case group) and 10 fertilized donors (as the control group) were analyzed for the volume of ejaculate and sperm concentration, motility and atypical morphology by computer-assisted semen analysis (CASA), the viability, chromatin structure and mitochondrial membrane potential (MMP) of the sperm stained by SYBR-14/PI, AO and JC-1 respectively, and assessed with flow cytometry. The results were analyzed through SAS software.

RESULTSU tests indicated that the semen from the infertile patients had not only lower concentration (U = 2.51, P = 0.0143), lower motility (U = 3.44, P = 0.001) and higher rate of atypical morphology (U = -5.88, P < 0.0001), but also lower viability (U = 4.72, P < 0.0001), MMP (U = -2.53, P = 0.0309), and chromatin integrity (alpha t: U = -3.82, P = 0.0003; SD alpha t: U = -3.98, P = 0.0001; COMP alpha t: U = -3.57, P = 0.0005). The multiple stepwise regression analysis revealed that sperm motility was positively correlated with sperm membrane integrity (t = 1.66, P = 0.1016), sperm MMP (t = 3.33, P = 0.0014) and sperm acrosome integrity (t = 3.24, P = 0.0019), while sperm MMP was negatively correlated with the rates of sperm neck and tail defects (t = -3.44, P = 0.001).

CONCLUSIONFlow cytometry plays a significant role in the evaluation of the quality of human sperm, and can be adopted as a useful tool in the diagnosis and treatment of male infertility.

Adult ; Case-Control Studies ; Cell Survival ; physiology ; Chromatin ; pathology ; Flow Cytometry ; Humans ; Infertility, Male ; physiopathology ; Male ; Membrane Potentials ; Middle Aged ; Sperm Motility ; Spermatozoa ; abnormalities ; pathology ; physiology

OBJECTIVETo investigate the clinical features and mutations of the FAH gene.

METHODClinical records of two cases were collected, and diagnosis was made according to the diagnostic criteria of the International Organization for Rare Disorders (NORD). Genomic DNA was extracted from peripheral blood leukocytes with QIAamp DNA Mini Kit. The DNA extracts were subjected to direct sequencing for 14 exons together with adjacent fragments of FAH gene using ABI Prism 3730 Genetic Analyzer (Applied Biosystems, Foster City, CA) after PCR based on genomic DNA. The mutation source was verified by analyzing parents' exons corresponding to patients' mutation exons. The homology between human FAH enzyme and that of other species was surveyed using software Clustal X(European Bioinformatics Institute, Hinxton, Saffron Walde, UK). Polyphen (Polymorphism Phenotyping), available online, were used to predict possible impact of an amino acid substitution on structure and function of FAH enzyme. Polyphen calculates position-specific independent counts (PISC) scores for two amino acid variants in polymorphic position. A PISC scores that differ by > 2 were regarded as indicating the probability of damaging variants.

RESULTPatient 1 was a 5 months and 21 days-old boy who suffered from persistent diarrhea, hepatomegaly, ascites; Alpha-fetoprotein > 1210 µg/L, levels of tyrosine in blood and succinylacetone in urine were 110.8 µmol/L and 83.7 µmol/L. His sister suffered from tyrosinemia type 1. Direct sequencing showed a G to A transition in CDS position 455 and 1027. He was compound heterozygous for the mutation c.455G > A/c.1027G > A, which predicts a change from tryptophan to a stop codon (TGG > TAG) at position 152 (W152X) and a change from glycine to arginine (GGG > AGG) at position 343 respectively. Patient 2 was a 6 year and 1 month-old girl with late-onset rickets who had signs of hepatosplenomegaly, rachitic rosary, windswept knees. Hypophosphatemia and alkaline phosphatase 1620 IU/L were detected. Alpha-fetoprotein 412.8 µg/L, levels of tyrosine in blood and succinylacetone in urine were 835.8 µmol/L and 27.48 µmol/L. Rickets did not improve after administration of calcium and vitamine D3. She is homozygous for the mutation c.1027G > A/c.1027G > A, which predicts G343R. The parents were mutation carriers. Analysis by Clustal X on the alignment of amino acids residual reservation among different species showed that the locative amino acid was highly conserved. Polyphen software predicted G343R was probably damaging (PISC score 3.235).

CONCLUSIONChildren with tyrosinemia type 1 can have manifestations of persistent diarrhea or late-onset rickets. Physical examination can reveal hepatosplenomegaly, laboratory tests indicate markedly elevated serum concentration of alpha-fetoprotein and alkaline phosphatase in plasma and succinylacetone in urine, other members in family may have tyrosinemias or parents are consanguineous. Mutations c.455G > A and c.1027G > A can be detected in FAH gene of Chinese children.

Amino Acid Sequence ; Base Sequence ; Child ; Child, Preschool ; DNA Mutational Analysis ; Diarrhea ; etiology ; genetics ; Exons ; Female ; Heptanoates ; urine ; Humans ; Hydrolases ; genetics ; Infant ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Rickets ; etiology ; genetics ; Tyrosine ; blood ; Tyrosinemias ; complications ; diagnosis ; genetics ; pathology ; alpha-Fetoproteins ; analysis

OBJECTIVETo investigate the plasma amino acid spectrum in infants more than 1-year-old with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) in order to identify potential diagnostic markers of NICCD.

METHODSInfants less than 1 year of age who had been referred to our hospital for investigation of suspected conjugated hyperbilirubinemia between June 2003 and June 2009 were eligible for enrollment. A total of 182 infants were enrolled and divided into three groups: infants diagnosed with NICCD (n = 24), according to gene testing and/or western blotting results; infants diagnosed with biliary atresia (BA; n = 20), according to intra-operative cholangiography findings; and infants diagnosed with idiopathic neonatal intrahepatic hepatitis (INH; n = 138), according to exclusionary findings for diseases affecting the extrahepatic biliary system and no positive serology results to indicate infections with hepatitis B, C, A or E, toxoplasmosis, rubella, herpes simplex, human immunodeficiency virus-1, or syphilis. The plasma amino acid spectrum of each infant was analyzed by tandem mass spectrometry (MS/MS). The concentrations of 18 amino acids, as well as the ratio of each amino acid to total amino acids, were compared among the three groups. Selected ratios of amino acids were analyzed by receiver operating characteristic (ROC) curve analysis.

RESULTSCompared with the BA and INH groups, the NICCD group had significantly lower levels of alanine (Ala; 175.7 and 205.7 vs. 136.3 mumol/L, P = 0.0001), aspartic acid (Asp; 47.5 and 43.1 vs. 31.55 mumol/L, P = 0.0041), glutamic acid (Glu; 276.16 and 263.24 vs. 175.71 mumol/L, P = 0.0075) and tryptophan (Trp; 41.90 and 47.97 vs. 28.51 mumol/L, P = 0.0003), but significantly higher levels of methionine (Met; 28.24 and 29.35 vs. 71.40 mumol/L, P = 0.0390), tyrosine (Tyr; 55.8 and 57.02 vs. 116.81 mumol/L, P = 0.0072) and citrulline (Cit; 15.09 and 15.65 vs. 97.42 mumol/L, P = 0.0001). The ratio of each amino acid to total amino acids showed the same trends for the NICCD group. The calculated areas under the ROC curves of the ratios of Cit, Tyr, and Met to the total amino acids were 0.874 (95% CI: 0.752 - 0.996), 0.814 (95% CI: 0.706 - 0.923), and 0.705 (95% CI: 0.535 - 0.875) respectively. The calculated area under the ROC curve of the ratio of Cit to Ala was 0.893 (95% CI: 0.781 - 1.005), and when the cut-off value of the ratio of Cit to Ala was 0.14 for diagnosis of NICCD, the sensitivity and specificity were 75% and 95% respectively.

CONCLUSIONThe plasma amino acid spectrum may represent a diagnostic indicator for NICCD, particularly the ratio of Cit to Ala.

Amino Acids ; blood ; Citrullinemia ; blood ; Female ; Humans ; Infant ; Male ; Tandem Mass Spectrometry

OBJECTIVETo detect the genes of Neisseria spp. isolated from patients with male genitourinary tract infections, and to study the pathogenicity of non-gonococcal strains of Neisseria and the laboratory diagnosis for the infections caused by Neisseria spp.

METHODSUsing polymerase chain reaction and nucleotide sequencing, we amplified and sequenced 4 genes of Neisseria spp. isolated from patients with male genitourinary tract infections, including 16S rRNA, orfl, cppB and nspA.

RESULTSFourteen Neisseria strains were identified through analysis of the 16S rRNA gene, including 3 N. mucosa strains, 3 N. cinerea strains, 2 N. gonorrhoea strains, 2 N. sicca strains, 2 N. subflava strains, 1 N. lactamica strain, and 1 N. polysaccharea strain. Among them, 9 showed positive results in gonococcal fluorescence-labeled multiplex-PCR detection, 1 in cppB gene reaction, 5 in orfl gene reaction, and 3 in nspA gene reaction. The consistency rate was 85.7% between the above results from our gene detection and those from the routine bacteriological methods.

CONCLUSIONThe cppB gene is absent in the non-gonococcal strains of Neisseria spp. that can cause male genitourinary tract infection. Most of the strains not only lack virulence-associated orfl and nspA genes, but also show positive results in gonococcal fluorescence-labeled multiplex-PCR detection, which is one of the important reasons for the misdiagnosis and missed diagnosis of gonorrhea infection. The combination of routine bacteriological methods and gene detection in laboratory examinations may help improve the accuracy rates of Neisseria species identification and clinical diagnosis of the infections caused by Neisseria spp.

Genes, Bacterial ; Gonorrhea ; diagnosis ; microbiology ; Humans ; Male ; Neisseria gonorrhoeae ; classification ; genetics ; Polymerase Chain Reaction ; Urinary Tract Infections ; microbiology

OBJECTIVETo investigate the expression of VEGF-C and VEGF-D and their correlations with lymphangiogenesis and angiogenesis in gallbladder carcinoma.

METHODSFifty cases of gallbladder carcinoma with complete clinical and pathological data were analyzed. The expression of VEGF-C and -D, D2-40, CD31 was assayed by immunohistochemical staining, with 10 samples of normal gallbladder tissues away from cancer and 19 samples of chronic cholecystitis as controls, and their correlation with clinicopathological findings were analyzed retrospectively.

RESULTSThirty-two (64.0%) of the 50 gallbladder cancers were positive for VEGF-C protein expression by immunohistochemistry and the positive rate of VEGF-D protein expression was 62.0% (31/50). The protein expression of VEGF-C and VEGF-D in tumor tissues was significantly higher than that in normal gallbladder tissues away from the tumor (P < 0.05), but no correlation with that in chronic cholecystitis (P < 0.05). The VEGF-C expression correlated with the patient age and lymph node metastasis (both P < 0.05). The VEGF-D expression only correlated with lymph node metastasis (P < 0.05). In the 50 gallbladder cancers, the MLVD was 6.9 + or - 3.6 and the MVD was 36.1 + or - 12.8. The MLVD in both VEGF-C and -D positive groups was significantly higher than that in the negative groups (P = 0.000), and the lymph node metastasis also increased. MVD in both VEGF-C and -D positive groups was higher than that in the negative groups (P < 0.05), and it was also correlated with tumor differentiation (P < 0.05). A significant positive correlation was also found between VEGF-C and VEGF-D expression (r = 0.498, P < 0.01).

CONCLUSIONVEGF-C and VEGF-D are involved in the lymphangiogenesis and angiogenesis in gallbladder carcinoma, promote lymph node metastasis of the tumor, and both are important in the regulation of lymphangiogenesis and angiogenesis in this cancer. VEGF-C and VEGF-D are of clinical significance in evaluating lymph node metastatic potency and estimation of prognosis in gallbladder carcinoma.

Adenocarcinoma ; blood supply ; metabolism ; pathology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Carcinoma, Papillary ; blood supply ; metabolism ; pathology ; Cholecystitis ; metabolism ; pathology ; Female ; Gallbladder Neoplasms ; blood supply ; metabolism ; pathology ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; Retrospective Studies ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

BACKGROUNDRoutine anteroposterior radiographs of the acromioclavicular (AC) joint with or without weight bearing have limitations in demonstrating the AC joint. Transarticular fixation with Kirschner wire is a treatment choice for AC dislocations. However, percutaneous fixation of the AC joint is technically demanding. The C-arm fluoroscopy can be used as routine intraoperative guidance to facilitate this procedure. The current study aims to introduce new projections, the axial and tangential views of AC joint, to help evaluate the severity of the injury and facilitate the percutaneous procedure.

METHODSThree shoulder specimens were used to find the projection directions of the axial and tangential views of the AC joint by using the digital radiography (DR) unit. The axial and tangential views were taken of 20 adult volunteers by referencing the projection directions determined in the shoulder specimens. The angles showed on the DR system and the angles between the coronal plane of the body and the vertical plane of the flat panel detector (FPD) during taking these radiographs were recorded. The C-arm fluoroscopy unit was used to take the axial and tangential views referencing the angles measured on the DR system. Routine anteroposterior radiographs of the AC joint were taken on the volunteers. The minimal distances from the distal clavicle to the acromion were measured on both tangential and anteroposterior radiographs. The data was statistically analyzed.

RESULTSThe clear axial and tangential radiographs of AC joints of the volunteers were obtained using both DR and C-arm fluoroscopy units. The angles demonstrated on the DR window are (20.8 ± 2.4)° for male and (18.3 ± 2.3)° for female. During taking the axial views, the angles between the coronal plane of the body and vertical plane of FPD are (23.3 ± 3.2)° for male and (20.1 ± 2.4)° for female. During taking tangential views, the corresponding angles are (117.5 ± 3.7)° for male and (113.1 ± 3.3)° for female. On the tangential radiographs, the minimal distance from the distal clavicle to the acromion is (6.1 ± 1.2) mm, wider than the same measurement on the anteroposterior radiographs (P < 0.05). Statistical analyses showed no significant differences in the above-mentioned angles and the minimal distances between the left and right AC joints (P > 0.05). There were no significant differences in the above-mentioned angles between DR and C-arm fluoroscopy units (P > 0.05).

CONCLUSIONSThe axial and tangential radiographs of the AC joint can demonstrate the joint clearly and they can be easily obtained with both DR system and C-arm fluoroscopy unit in similar projection directions.

Acromioclavicular Joint ; diagnostic imaging ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Radiographic Image Enhancement

Objective Toinvestigatethestatusoftick-borneRickettsiaeinfectionsamongmurine-likeanimalsin differentareasofZhejiangprovince.Methods Liverandspleensamplesofmurine-likeanimalscapturedthroughnight trapping method were collected from Anji,Jinhua and Tiantai County according to their geographic locations and historical detection of Rickettsiae .Nest-PCR tests were used to determine the presence of the 16S rRNA genes of Anaplasma and Ehrlichia ,and the heat shock protein genes (groEL)of Rickettsiae (including typhus and spotted fever group)and Orientiainthesesamples.Results Atotalof851murine-likeanimalsbelongingto14specieswerecaptured.The predominant species were Rattus confucianus (30.32%),Apodemus agrarius (18.80%) and Thallomys paedulcus (1 1.75%)and they were significantly different among three areas (P<0.05 ).48 Rickettsia positive were found in 562 tested samples with the positive rate of 8.54%,among which the percentage of Anaplasma,typhus group Rickettsia, Orientia,Ehrlichia and spotted fever group Rickettsia were 3.38%,1.78%,1.78%,1.07% and 0.53% respectively. The positive rates of Anaplasma in Jindong (4.76%)and Anji (4.27%)were significantly higher than that in Tiantai (P<0.05 )while the spotted fever group Rickettsia were found only in Tiantai County.Moreover,Rattus confucianus-the predominant species of Zhejiang Province-had the highest infection rate of tick-borne Rickettsiae up to 14.97%.Co-infections with several Rickettsiae were existed among the same species.Conclusion Rickettsiae infections exist widely among different areas of Zhejiang province and the positive rates are significantly different among species.

OBJECTIVEThis study was designed to compare clinical efficacy of segmental pulmonary vein ablation (SPVI), amiodarone or amiodarone plus losartan on sinus rhythm maintenance in patients with lone paroxysmal atrial fibrillation (PAF).

METHODSPatients with lone PAF were treated with amiodarone alone (A, n = 52), segmental pulmonary vein isolation (SPVI, n = 51), or amiodarone plus losartan (AL, n = 51). The primary endpoint of this study was the incidence of symptomatic atrial tachyarrhythmia (> 30 s) documented by 12 lead ECG or Holter during 12 months follow-up period.

RESULTSDuring follow-up, AF was documented in 24 patients (46.2%) in A group, 11 patients (21.6%) in SPVI group and 12 (23.5%) in AL group (P < 0.05 vs. A group). The Kaplan-Meier survival analysis demonstrated a significant equally reduction in AF recurrence in SPVI and AL groups (P = 0.009, log-rank test and P = 0.018, log-rank test, respectively) compared with A group. The hazard ratio for AF recurrence in patients treated with SPVI and amiodarone plus losartan was 0.41 (95% CI 0.200 to 0.848, P = 0.016) and 0.46 (95% CI 0.225 to 0.953, P = 0.036), respectively. Incidences of major adverse cardiac events were similar among the groups (9.6% in A, 3.9% in SPVI and 7.8% in AL group, P > 0.05).

CONCLUSIONThe results of this study suggest that the segmental pulmonary vein isolation and amiodarone plus losartan are superior to amiodarone alone for preventing AF recurrence in patients with lone PAF.

Aged ; Amiodarone ; therapeutic use ; Anti-Arrhythmia Agents ; therapeutic use ; Atrial Fibrillation ; prevention & control ; therapy ; Catheter Ablation ; methods ; Combined Modality Therapy ; Follow-Up Studies ; Humans ; Losartan ; therapeutic use ; Middle Aged ; Prospective Studies ; Treatment Outcome