Objective To determine whether arsenic has estrogen-like effects,the cell proliferation was measured iil human eervical cancer line(HeLa)in vitro.Methods The HeLa cells were grown in improved RPMI 1640 supplemented respectively with β-estradiol(E2,1 nmol/L),Arsenic trioxide(As2O3,0.5,1.0,5.0 μmol/L),ICI (500 nmol/L),E2(1 nmol/L)+ICI(500 nmol/L),As2O3(1.0 μmol/L)+ICI(500 nmol/L)and control.The growth morphology of HeLa cell was observed under microscope after 72 h.The method of M1Tr was used to study the cell proliferation after 24.48 and 72 h.The technique of flow eytometry was used to measure cell cycle after 48 h. Results HeLa cells in E2 and 0.5 μmoL/L As2O3 treatment were more better growth in morphology than control group.Percentage of HeLa cells proliferation at 24,48,72 h in E2 and 0.5 μmol/L As2O3 treatment were 6.35%, 11.56%,38.33%and 6.35%,8.50%,20.26%respectively.The proliferation effect of HeLa cells was similar in two treatments.The proliferation of HeLa cells were inhibited in other treatments.Compared with control[(41.68± 1.05)%],HeLa cells were promoted go to S phases in E2[(55.72±2.31)%]and 0.5 μmol/L As2O3[(47.82± 1.41)%]treatment.But in other treatments HeLa cells were hold back to S phases.Compared with control,there was a significant differenee(P＜0.05)of cell percentage in S phases in 5.0 μmol/L As2O3[(21.11±4.99)%]and ICI[(20.16±4.76)%]treatments.Conclusion Small amounts of As2O3 impose estrogen.1ike effects and stimulate the proliferation of HeLa cells.

Day-old chick is unique animal model in brain development and behavior study. The intermediate medial mesopallium (IMM), a region of the chick forebrain, is intimately involved in the early learning processes, which offers the ideal opportunity to study the neural changes that underlie behavioral plasticity. In this paper, the intracellular recordings were conducted from IMM neurons in chick forebrain slices, in which electrophysiological properties, synaptic responses and long-term potentiation (LTP) were observed. Coronal sections of left forebrains (500 mum thick), containing IMM, were prepared from domestic chicks, aged 2-10 days. In 69 IMM neurons, the resting membrane potential was measured to be (-59.4+/-5.3) mV, slope membrane resistance (70.8+/-27.2) MΩ, and time constant (10.2+/-4.3) ms. The amplitude, threshold, overshoot, half-width, max rise slope and max decay slope of action potential evoked by intracellular current injection were (85.2+/-9.4) mV, (-38.7+/-7.6) mV, (25.6+/-8.9) mV, (2.1+/-0.5) ms, (150.5+/-41.2) mV/ms and (-64.3+/-14.0) mV/ms, respectively. Spike-firing frequency was increased with depolarizing current intensity in 32 of 69 tested cells [linear regression slope was (21.5+/-10.9) Hz/nA, P<0.05 in all cells]. The depolarizing synaptic responses (i.e. EPSPs), with stimulus intensity- and membrane potential-dependent properties, were elicited by dorsal (n=25) or ventral (n=62) focal electrical stimuli at 0.1 Hz in all tested IMM neurons and could be nullified reversibly by perfusion with 100 mumol/L AP5 (NMDA receptor antagonist) and 3 mumol/L DNQX (non-NMDA receptor antagonist), but enlarged by 6 mumol/L bicuculline (GABA(A) receptor antagonist). The EPSPs evoked by ventral stimulation were persistently increased after tetanic stimulation (5 Hz, 300 pulses/train, 2 trains, train interval 10 min) in 6 of 12 tested IMM neurons. The amplitude of EPSPs was potentiated to more than 120% of control level (when analyzed at 45 min of enhancement, P<0.05, n=5), which lasted at least 30 min and then could be referred to as LTP. Moreover, area under curve, duration and max rise slope of EPSPs were also enhanced (P<0.05), while no significant changes were observed in the electrophysiological parameters of IMM neurons following induction of LTP (P>0.05). These results suggest that the intracellular recording techniques in the chick brain slices can be used to perform multi-parameter analysis of synaptic responses and their LTP.
Animals ; Brain ; physiology ; Chickens ; In Vitro Techniques ; Long-Term Potentiation ; Membrane Potentials ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors

OBJECTIVETo investigate the relationship between C-reactive protein (CRP) + 1444C/T, CRP+1059G/C polymorphisms and chronic periodontitis (CP) in a Han Chinese population.

METHODSClinical periodontal parameters [attachment loss (AL) probing depth (PD) and bleeding on probing (BOP)], and serum CRP levels were examined in CP patients (n = 126) and healthy subjects (n = 113).

RESULTSThe mean serum CRP level [(1.74 ± 1.67) mg/L] was significantly higher in the CP group than in the control group [(0.57 ± 0.39) mg/L], P < 0.001. In the control group, serum CRP levels were significantly lower in subjects with the CRP +1059 GC and CC genotypes than those with the CRP +1059 GG genotype (P < 0.01). There was no significant difference between genotypes in the CP group. In CP and the control groups, serum CRP levels were significantly higher in subjects with the CRP + 1444 CT and TT genotypes compared to those with the CRP + 1444 CC genotype (P < 0.5). The percentage of CRP + 1059 C allele was 6.7% (17/252) in the CP group and 4.9% (11/226) in the control group. The percentage of CRP + 1444 T allele was 6.3% (16/252) in the CP group and 5.3% (12/226) in the control group (P > 0.5). There was no significant difference between groups in both allele frequencies (P > 0.5). The association of CRP + 1059G/C, CRP + 1444 C/T polymorphisms with CP was not found in a regression model (P > 0.5).

CONCLUSIONSThe presence of a CRP + 1059C-allele was associated with lower serum CRP levels and the presence of a CRP + 1444T-allele was associated with higher serum CRP levels. However, the data suggested that CRP + 1059G/C, CRP + 1444 C/T polymorphisms were not significantly associated with serum CRP levels of chronic periodontitis patients in ethnic Han Chinese.

Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; C-Reactive Protein ; genetics ; Chronic Periodontitis ; complications ; genetics ; metabolism ; Coronary Disease ; complications ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Periodontal Index ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the types of disc position of the temporomandibular joint (TMJ) and their relationship with age and gender in asymptomatic volunteers by magnetic resonance imaging (MRI).

METHODSA total of 100 asymptomatic volunteers were divided into 5 groups (11-, 21-, 31-, 41-, 51 - 60 years old), 10 male and 10 female were included in each group. A total of 200 TMJs were scanned with Siemens Trio Tim 3.0T MRI system oblique-sagittally at closed- and open-mouth positions. Visual evaluation of MR images were performed on 3 neighbour slices of each TMJ (lateral, central, medial).

RESULTSThere was a normal disc-condyle relationship in 59 of the 100 asymptomatic subjects. The normal disc position, anterior disc displacement and insidious anterior disc displacement (IADD) were observed in 140 (70.0%), 14(7.0%) and 46(23.0%)joints respectively, which showed no significant differences among different age groups or between genders (P > 0.05). The maximum mouth opening of the 100 asymptomatic volunteers was (46.3 +/- 5.5) mm, with no significant difference among the various types of disc position (P > 0.05).

CONCLUSIONSDisc displacement does exist in the asymptomatic volunteers, however, its occurrence is not associated with age and gender. IADD is the main type of the disc displacement in asymptomatic volunteers. There is no close relationship between the imaging manifestation of the disc displacement and clinical symptoms.

Adolescent ; Adult ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Temporomandibular Joint ; anatomy & histology ; pathology ; Temporomandibular Joint Disorders ; pathology

OBJECTIVETo investigate the clinical treatment modality and prognosis of small cell lung cancer(SCLC).

METHODWe retrospectively analyzed the clinical data of 77 SCLC patients who were admitted to our department after 2002.

RESULTSThe disease was limited in 43 patients and extensive in 34 patients. For patients with limited SCLC, the 1-year, 2-year, and 5-year survival rate was 80%, 56%, and 21%, respectively. Four patients who had undergone surgical resection were all alive. Among patients who underwent adjuvant chemotherapy followed by radiotherapy, salvage chemotherapy, and salvage chemotherapy followed by radiotherapy, the median of survival period was 51 months, 12 months, and 28 months, respectively. For patients with extensive SCLC, the 1-year and 2-year survival rate was 56% and 25%, respectively. The median of survival period was 14.3 months. Stage was an independent factor in multifactor COX regression. Monofactor COX regression showed that radiotherapy and resection were factors correlated with survival. Brain metastasis had no impact on survival.

CONCLUSIONSChemotherapy followed by radiotherapy is preferred for limited SCLC, while surgical resection remains questionable for early-stage patients. For extensive SCLC, multi-line chemotherapy may be helpful to improve the overall survival. Stage is an independent factor for predicting the prognosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Small Cell ; diagnosis ; therapy ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; diagnosis ; therapy ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Analysis

OBJECTIVETo analyze factors influencing prognosis and to develop a prognosis predicting model for patients with chronic severe hepatitis.

METHODSClinical data from 408 patients suffering from chronic severe hepatitis were divided into improved group and deteriorated group. The main clinical and laboratory variables were analyzed as predictive factors of prognosis with logistic regression analysis.

RESULTSIt was shown that age, sex, TBil, DBil, [Cl-], [Na+], WBC, MCV, PT, NH3, PTA, and BUN were different between the two groups with statistical significance (P < 0.05). The prognosis predicting model was P = 1/(1 + e(-y)), Y = -4.636 + 0.022X1 + 0.034X2 + 0.096X3 + 0.047X4 - 0.042X5, (X1-age, X2-TBil, X3-BUN, X4-MCV, X5-PTA).

CONCLUSIONAge, TBil, BUN, MCV and PTA are the independent risk factors related to prognosis of chronic severe hepatitis.

Adult ; Female ; Hepatitis, Chronic ; diagnosis ; physiopathology ; Humans ; Logistic Models ; Male ; Middle Aged ; Prognosis ; Risk Factors

Objective Little is known about the effect of RNAi on mitochondrial apoptotic pathways. This study aims to explore the effects of the Survivin shRNA-APC double-gene on colon cancer mitochondrial apoptosis pathway-related factors survivin,cytochrome C (Cytc),second mitochondria-derived activator of caspases (Smac),and cysteine aspartic acid specific protease 9 (Caspase-9) as well as on the apoptosis of colon cancer transplanted tumor (CCTT) cells. Methods Thirty nude mice were randomly divided into five groups of equal number,Survivin shRNA-APC double-gene,survivin shRNA,APC,empty vector and blank transfection. The CCTT model was established in the nude mice by subcutaneous injection of the colon cancer cell strains stably transfected with the Survivin shRNA-APC double-gene,survivin shRNA,APC,an empty vector and HT-29,respectively,into the mid-posterior part of the left armpit of the nude mice. The rate of tumor growth inhibition was calculated by measuring the volume and weight of the CCTTs in the nude mice. The mRNA and protein expressions of survivin,Cytc,Smac and Caspase-9 in the tumor tissue were detected by real time PCR and immunohistochemistry,respectively,and the apoptosis rate of the CCTT cells was detected by TUNEL. Results The model of CCTT was successfully established in the nude mice. Com-pared with the empty vector and blank transfection groups,the mice in the double-gene,survivin shRNA and APC groups showed sig-nificantly decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). In comparison with the survivin shRNA and APC groups,the double-gene group exhibited remarkably decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). The mRNA and pro-tein expressions of survivin were significantly lower while those of Cytc,Smac and Caspase-9 markedly higher in the double-gene,sur-vivin shRNA and APC groups than in the empty vector and blank transfection groups (P<0.05),the former even lower (P<0.05) and the latter even higher in the double-gene than in the survivin shRNA and APC groups (P<0.05). The apoptosis rate of the CCTT cells was significantly increased in the double-gene ([56.78±3.04]%),survivin shRNA ([33.61±2.02]%) and APC groups ([30.16± 1.72]%) as compared with the empty vector ([10.05±0.42]%) and blank transfection groups ([9.87±0.30])% (P<0.05),even higher in the double-gene group than in the survivin shRNA and APC groups (P<0.05). Conclusion The Survivin shRNA-APC double-gene may induce apoptosis of colon cancer transplanted tumor cells by down-regulating the expression of the apoptosis inhibitor survivin,upregulating the expressions of Cytc,Smac and Caspase-9,and suppressing the growth of the colon transplanted tumor,with more significant abilities than a single gene in regulating apoptosis-related factors,inducing cell apoptosis and inhibiting the growth of the transplanted tumor.

This study was aimed to evaluate the long-term effects of telbivudine (LdT) in the treatment of chronic hepatitis B (CHB) and HBV-related liver cirrhosis (LC) and to observe the changes of immunological responses during LdT treatment. Clinical data of 80 CHB and 28 HBV-related LC patients who were administered with LdT for 108 weeks and followed up were retrospectively analyzed. The liver function indicators including ALT, AST and γ-GT, HBV DNA copy number in serum and the rates of hepatitis B e antigen (HBeAg) seroconversion were analyzed before and 12, 24, 36, 48, 60, 72, 84, 96 and 108 weeks after LdT treatment in CHB and LC groups. Four serum fibrosis-related markers, including hyaluronic acid (HA), human laminin (LN), human type IV collagen (IV-C) and human N-terminal procollagen III peptide (PC-III), were detected before and after LdT treatment in LC group. The results showed favorable viral suppression and biochemical responses after treatment with LdT for 12 weeks, and a high rate of virological and biochemical control was maintained during the course of 108-week treatment in both CHB and LC groups. The four fibrosis-related markers, especially HA and LN, were down-regulated to some degrees in LC group. Moreover, LdT treatment led to the fluctuation of the circulating interferon-γ (IFN-γ) and interleukin-10 (IL-10) levels at different time points in CHB group. It was concluded that LdT could favorably lead to the virological suppression and biochemical remission. Besides, IFN-γ and IL-10 may represent a suitable and effective predictor of responsiveness during LdT therapy.
Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Liver Cirrhosis ; drug therapy ; immunology ; Male ; Middle Aged ; Thymidine ; analogs & derivatives ; therapeutic use

An infectious cDNA clone of H120 vaccine strain of infectious bronchitis virus (IBV) was constructed to demonstrate its potential as a gene transfer vector. Primers were designed according to the published genome sequence of H120 strain, and ten cDNA fragments covering the entire genome of H120 strain was amplified by RT-PCR. All the PCR products were ligated into pMD19-T vector and sequenced, and the ORF5a open reading frame in the pMDTM9 plasmid was replaced by an enhanced green fluorescent protein (EGFP) gene. Recombinant plasmids were digested by the restriction enzyme Bsa I, and all the cDNA fragments were recovered. By using appropriate ligation strategy, the genomic cDNA of H120 strain were reconstituted. Then genome RNA was synthesized in vitro by T7 RNA polymerase and transfected into BHK-21 cells. Recombinant virus expressing the green fluorescent protein was rescued and identified by RT-PCR and sequencing. The characteristics of recombinant virus were evaluated by passage in embryonated chicken eggs. This study showed that the 5a ORF is a good candidate for an insertion site of recombinant genes for the development of IBV infectious cDNA clone as a gene transfer vector.
Animals ; Cell Line ; Chick Embryo ; Cricetinae ; DNA, Complementary ; genetics ; metabolism ; Gene Expression ; Gene Transfer Techniques ; instrumentation ; Genetic Vectors ; genetics ; physiology ; Green Fluorescent Proteins ; genetics ; metabolism ; Infectious bronchitis virus ; genetics ; physiology

Objective To investigate the effects of different dialysates on expression of protein kinase C-? (PKC?) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKC? specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKC? mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group, normal control group and fluid A+powder B+rottlerin group (P0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKC?. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.