The conductivity and permittivity of blood in mice were measured by the AC electrical impedance method at frequency range of 0.1-100MHz, and then the changes of the Cole-Cole parameters of dielectric spectra of blood from phenylhydrazine-induced anemia mice were observed by numerical calculation and curve fitting residual analysis of the Cole-Cole equation. The results showed that hematocrit (Hct) of the mice with phenylhydrazine injection was significantly reduced; the permittivity(epsilon) spectroscopy of blood moved to the low insulating region and its permittivity decreased; conductivity (kappa) spectrum curve of blood moved to the high conductivity zone and conductivity increased; the 2nd characteristic frequency was lower than that in the normal group. There was phenylhydrazine dose dependent in the changes of the Cole-Cole parameters of dielectric spectra of blood.
Anemia ; blood ; Animals ; Blood Physiological Phenomena ; Electric Conductivity ; Electric Impedance ; Hematocrit ; Mice

BACKGROUNDDiabetic macrovascular complications are important causes of cardiovascular and cerebrovascular diseases and also one of the major causes of morbidity and mortality in patients with type 2 diabetes mellitus (T2DM). Phlorizin has been reported to be effective in reducing the blood glucose level in diabetic mellitus, while little is known about its effects on vascular complications. This study aimed to observe the effects of phlorizin on the aorta of diabetes db/db mice and explore its mechanism.

METHODSDiabetic db/db mice (n = 16) and age-matched db/m mice (n = 8) were divided into three groups: normal control group (CC group, db/m mice, n = 8), untreated diabetic group (DM group, db/db mice, n = 8) and diabetic group treated by phlorizin (DMT group, db/db mice, n = 8). Phlorizin (20 mg/kg body weight) was given in normal saline solution intragastrically for 10 weeks. Animals were weighed weekly. At the 10th weekend, all mice were fasted overnight and then sacrificed. Fasting blood was collected, and the aortas were dissected. The blood samples were analyzed for fasting blood glucose (FBG), serum advanced glycation end products (AGEs), malondialdehyde (MDA) and superoxide dismutase (SOD) activity, the aortic ultrastructure was studied.

RESULTSThe weight and serum concentration of FBG, AGEs, and MDA in the DM group were higher than that in the CC group (P < 0.01), and they were significantly lower in the DMT group (P < 0.05). Serum SOD activity was lower than that in the CC group (P < 0.01), and it is significantly higher in the DMT group (P < 0.05). The severity of aorta damage in the DMT group was less than that in the DM group.

CONCLUSIONSPhlorizin protected the db/db mice from diabetic macrovascular complications, attributed to the decreasing of blood glucose and AGEs level, and its antioxidant potential. This study may provide a new natural medicine for treating diabetic macrovascular complications.

Animals ; Aorta, Thoracic ; pathology ; Blood Glucose ; analysis ; Diabetic Angiopathies ; drug therapy ; pathology ; Glycation End Products, Advanced ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phlorhizin ; therapeutic use ; Superoxide Dismutase ; metabolism

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

OBJECTIVETo analyze the expression and clinical values of HPV L1 capsid protein and p16INK4a protein in uterine cervical lesions.

METHODSFifty-four cervical intraepithelial neoplasias CIN1, 44 CIN2, 78 CIN3, and 48 squamous cell carcinoma were included in this study. All CIN and squamous carcinomas were stained with anti-HPV L1 capsid protein antibodies and anti-p16INK4a antibody. Forty-five CIN1 patients were followed up for 6 years.

RESULTSForty-five CIN1 patients were followed up for 6 years, among them 6 cases showed a progression (One case changed to CIN3, 5 cases to CIN2). L1 positivity was found in 50 cases which decreased with CIN increasing (χ(2) = 259.923, P < 0.001) while p16INK4a positivity was found in 177 cases which co-increased with CIN (χ(2) = 48.842, P < 0.001). L1(-)p16INK4a (-) or L1(+)p16INK4a(-) appeared mainly in CIN1 while L1(-)p16INK4a(+) appeared mainly in CIN2 lesions. No progression was found in the group of L1(-)p16INK4a(-) CIN1 patients. The risk of CIN1 progression in L1(-)p16INK4a(+) group was 66.7% while L1(+)p16INK4a(-) group was 9.5%, and L1(+)p16INK4a(+) group was 33.3%.

CONCLUSIONSThe expression of p16INK4a together with HPV L1 are different in various cervical lesions, and the combined detection of p16INK4a and HPV L1 can be helpful for estimating the biological potentiality of CIN lesions.

Adult ; Aged ; Capsid Proteins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; virology ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Disease Progression ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Oncogene Proteins, Viral ; metabolism ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; metabolism ; pathology ; virology ; Young Adult

This review presents the state of the art of pH-responsive polymeric micelles for cancer drug delivery. Solid tumors have a weakly acidic extracellular pH (pH < 7), and cancer cells have even more acidic pH in endosomes and lysosomes (pH 4-6). The pH-gradients in tumor can be explored for tumor targeting and drug release in cancer drug delivery by applying pH-responsive polymeric micelles. The pH-responsive polymeric micelles consist of a corona and a core, and are made of amphiphilic copolymers, in which there are pH-responsive polymeric blocks. Two types of pH-responsive polymers-protonizable polymers and acid-labile polymers have been mainly used to make pH-responsive micelles for drug delivery. The protonizable polymers are polybases or polyacids, and their water-soluble/insoluble or charge states undergo changes with the protonation or deprotonation stimulated by external acidity, while the acid-labile polymers change their physical properties by chemical reaction stimulated by the acidity. Polymeric micelles whose core or coronas respond to the tumor extracellular acidity can be explored for triggering the fast release of the carried drug, activating the targeting group and accelerating the endocytosis of drug-loaded polymeric micelles, and those whose core or coronas respond to the tumor lysosomal acidity can be used for facilitating their escape from the lysosomes and targeting the nucleus. Various in vivo and in vitro experiments demonstrated that pH-responsive polymeric micelles are effective for cellular targeting, internalization, fast drug release and nuclear localization, and hence enhancing the therapeutic efficacy and reducing the side effect of cancer chemical therapy.
Antineoplastic Agents ; administration & dosage ; therapeutic use ; Drug Delivery Systems ; Humans ; Hydrogen-Ion Concentration ; Micelles ; Nanoparticles ; Neoplasms ; drug therapy ; Polymers ; chemistry

OBJECTIVEProstate cancer (PCa) has the highest incidence among male malignancies in Western industrialized countries and, as a most common malignant disease in urology, its incidence has been increasing in recent years in Chinese men. This study was to investigate the risk loci associated with PCa susceptibility in Han Chinese by analyzing single nucleotide polymorphisms (SNP).

METHODSWe collected peripheral blood samples from 1 667 PCa patients and 1 525 healthy men, and detected 40 loci associated with PCa susceptibility by analyzing SNPs using Sequenom technology.

RESULTSOf the 40 known loci, 16 were confirmed to be significantly associated with PCa susceptibility (P < 0.05). The loci 1, 2 and 5 at 8q24, 10q11 and 22q13.2 also contributed to PCa susceptibility in different ethnic groups.

CONCLUSIONPCa susceptibility is obviously associated with the risk loci rs1465618, rs721048, rs12621278, rs7679673, rs12653946, rs339331, rs1512268, rs10086908, rs16901979, rs1447295, rs10993994, rs10896449, rs902774, rs9600079, rs11649743 and rs5759167 in Chinese Han population.

Aged ; Asian Continental Ancestry Group ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk Factors

OBJECTIVETo investigate the effects of a Chinese herbal extract Songyou Yin on residual hepatocellular carcinoma after chemotherapy in nude mice and the relevant mechanisms.

METHODSOrthotopic nude mouse models bearing residual hepatocellular carcinoma after chemotherapy was established using human liver carcinoma MHCC97L cells. Three different doses of Songyon Yin (2.1 g/kg, 4.2 g/kg and 8.4 g/kg) were administered to the mice in the trial groups by intragastric gavage, respectively. The mice in the control group were administered physiological saline. The tumor growth, metastasis and survival in the mice of each group were recorded. The corresponding mechanisms were studied.

RESULTSThe pulmonary metastasis rates of the control group and 2.1g/kg, 4.2g/kg, 8.4g/kg Songyou Yin treatment group were 86.7%, 73.3%, 40.0%, and 20.0%, respectively, and the survivals of these groups were 53.83 ± 4.71, 56.50 ± 6.09, 66.67 ± 5.61, 81.17 ± 7.36 days, respectively. Compared with the mice in the control group, mice in the 4.2 g/kg, 8.4 g/kg Songyou Yin treatment groups had a lower pulmonary metastasis rate (P = 0.021 and P = 0.001, respectively) and longer survival (P = 0.002 and P = 0.001, respectively). A restoration of E-cadherin expression and a concomitant reduction of N-cadherin expression were detected in the tumors of the 4.2 g/kg and 8.4 g/kg Songyou Yin treatment groups.

CONCLUSIONSSongyou Yin effectively inhibits the invasion and metastasis of the residual hepatocellular carcinoma after chemotherapy in nude mice through attenuating the epithelia-mesenchymal transition and prolongs the survival. Songyon Yin may have potential to promote the efficacy of chemotherapy in hepatocellular carcinoma.

Animals ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cadherins ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; Lung Neoplasms ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasm, Residual ; metabolism ; pathology ; Organoplatinum Compounds ; therapeutic use ; Plants, Medicinal ; chemistry ; Random Allocation ; Survival Rate ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

OBJECTIVETo determine whether Acanthamoeba polyphaga could affect the survival and growth of Vibrio cholerae O139 in low temperature.

METHODSV. cholerae O139 was co-cultured with the Acanthamoeba polyphaga to be examined on its intracellular growth and survival rate within cysts at low temperature, using methods as Gram-staining, electron microscope and passage culture.

RESULTSV. cholerae O139 were observed to enter into the trophozoites and grow the within the vacuoles after 8 hour incubation with Acanthamoeba polyphaga. The germs survived in the vacuole and/or endo-layer of wall and could be re-isolated from the excystment of Acanthamoeba polyphaga. At 30 degrees C, V. cholerae O139 could survive for 120 days with the amoeba while less than 45 days in PAS. At 4 degrees C, the number of viable bacteria decreased and reached undetectable levels for both study and control groups after a 30-day incubation. V. cholerae O139 could be re-isolated from the 30-, 45-, 60- and 75-day's infected cysts after excystment. However the ability of excystment for 90-day's infected cysts decreased and V. cholerae O139 within the cyst could not be isolated again because the amoebae had lysed.

CONCLUSIONThese findings indicated that V. cholerae O139 could grow within Acanthamoeba polyphaga and the survival time could be increased in the cysts at low temperature. It seemed that Acanthamoeba can provide an environmental reservoir for V. cholerae O139.

Acanthamoeba ; microbiology ; Bacterial Capsules ; Colony Count, Microbial ; Temperature ; Vibrio cholerae ; growth & development

OBJECTIVETo explore the factors that affect the recovery of consciousness in patients with disorders of consciousness following brain trauma.

METHODSWe analyzed the data of 114 patients with disorders of consciousness following brain trauma admitted for rehabilitation. Bilateral logistic regression analysis was used to explore the factors that affected the recovery of the patients' consciousness. A logistic regression model was established and the ROC curve was drawn to obtain the optimal threshold of the prognostic model.

RESULTSUnivariate analysis showed that vegetative state duration (P<0.001), CRS-R scores (P<0.001), hydrocephalus (P=0.037), hypertonia (P=0.034), central fever (P=0.035), paroxysmal sympathetic hyperactivity (PSH) (P=0.004), and epilepsy seizures were correlated with the recovery of consciousness. Logistic multivariate analysis showed that central fever (OR=3.493, P=0.044), vegetative state duration (OR=1.016, P=0.008), PSH (OR=4.223, P=0.034) and CRS-R scores (OR=0.640, P=0.002) all significantly affected the recovery of consciousness. The χvalue of the Hosmer-Lemeshow test was 10.214 (P=0.250), and the goodness of fit of this model indicated an outstanding fitting (c=0.91).

CONCLUSIONSThe presence of PSH is the one of the most important factor followed by centric fever to affect the outcome of patients with disorders of consciousness. A lower CRS-R score and a longer duration of vegetative state also predict a poor recovery of consciousness in these patients.

OBJECTIVETo investigate the expressions of costimulators on peripheral T and B lymphocytes in patients with idiopathic thrombocytopenic purpura (ITP).

METHODSThe expression of B7-CD(28) and CD(40) of peripheral lymphocytes was measured by flow cytometry in 21 ITP patients and 9 normal subjects. The expression of PAIgG was measured by ELISA method.

RESULTSThe expression of CD(4)(+)CD(28)(+) was lower in ITP patients than in normal controls, but the expression of CD(86)(+) and CD(86)(+)CD(19)(+) was higher in ITP patients than in normal controls, while the expression of CD(80)(+), CD(40)(+), CD(28)(+), CD(19)(+)CD(86)(+), CD(19)(+)CD(40)(+), CD(4)(+)CD(28)(+)/CD(4)(+), CD(19)(+)CD(80)(+)/CD(19)(+) and CD(19)(+)CD(40)(+)/CD(19)(+) in ITP patients was normal. The PAIgG level was higher in 16 patients with a mean of (184.62 +/- 38.00) ng/10(7) plt. A positive correlation was found between PAIgG and CD(19)(+)CD(86)(+)/CD(19)(+) expression.

CONCLUSIONThere is no deficiency in expression of CD(28) on CD(4)(+) T lymphocytes in ITP patients. The change of CD(86) expression on B lymphocytes is possibly involved in pathophysiology of ITP, which may provide a theoretical instruction for ITP patients immunological therapy.

Adolescent ; Adult ; Antigens, CD ; blood ; Blood Platelets ; immunology ; Female ; Humans ; Immunoglobulin G ; blood ; Lymphocyte Activation ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood