Objective: To investigate the changes of the cell cycle regulators ATM, Chk2 and p53 and cell cycle arrest in HeLa cells after cisplatin therapy. Methods: The proliferation-inhibiting rates of HeLa cells induced by cisplatin of different concentrations were measured by MTT assays. The mRNA and protein expressions of ATM, Chk2 and p53 of HeLa cells with and without cisplatin were detected by RT-PCR and Western blot, respectively. The cell cycle analysis was conducted by flow cytometric analysis. Results: Cisplatin inhibited the proliferation of HeLa cells in a dose- and time-dependent manner. The mRNA and protein expressions of ATM, Chk2 and p53 were increased in HeLa cells treated with cisplatin. The cell cycle was arrested in G2/M phase in HeLa cells treated with cisplatin. Conclusion: Activation of ATM, Chk2 and p53 might be critical in determining whether cells survive or undergo apoptosis. Targeting ATM, Chk2 and p53 pathway might be a promising strategy for reversing chemoresistance to cisplatin in cervical cancer.

Objective: To investigate the changes of the cell cycle regulators ATM, Chk2 and p53 and cell cycle arrest in HeLa cells after cisplatin therapy. Methods: The proliferation-inhibiting rates of HeLa cells induced by cisplatin of different concentrations were measured by MTT assays. The mRNA and protein expressions of ATM, Chk2 and p53 of HeLa cells with and without cisplatin were detected by RT-PCR and Western blot, respectively. The cell cycle analysis was conducted by flow cytometric analysis. Results: Cisplatin inhibited the proliferation of HeLa cells in a dose- and time-dependent manner. The mRNA and protein expressions of ATM, Chk2 and p53 were increased in HeLa cells treated with cisplatin. The cell cycle was arrested in G2/M phase in HeLa cells treated with cisplatin. Conclusion: Activation of ATM, Chk2 and p53 might be critical in determining whether cells survive or undergo apoptosis. Targeting ATM, Chk2 and p53 pathway might be a promising strategy for reversing chemoresistance to cisplatin in cervical cancer.

AIM:To compare the refractive errors measured by the VISX WaveScan, OPD - Scan Ⅲ and the subjective refraction.
METHODS: Seventy - six patients ( 152 eyes ) were recruited from January 2013 to December 2013. All patients were measured with subjective refraction by the phoropter (NIDEK, RT-5100), objective refraction by the WaveScan ( AMO Company, USA) , OPD-ScanⅢ ( Nidek Technologies, Japan). The sphere, cylinder, axis of the three methods were compared and analyzed.
RESULTS: The sphere measured by WaveScan was lower than that by subjective refraction, the difference was 0. 13±0. 30D (t=3. 753, P<0. 001). For cylinder, the difference was 0. 13±0. 43D (t=3. 664, P<0. 001). There was no significance for sphere, cylinder, and spherical equivalent between OPD - Scan Ⅲ and subjective refraction (P>0. 05). The value of the difference between WaveScan and subjective refraction was 5. 87o±6. 19o for the axis and the difference between OPD-Scan Ⅲ and subjective refraction was 3. 82o±3. 95o. There was statistic significance (t=2. 817, P=0. 006).
CONCLUSION: For sphere and cylinder, WaveScan generated some deviation relative to subjective refraction. The Nidek OPD-ScanⅢ gives more accurate measures of objective refraction when compared with subjective refraction.

OBJECTIVETo study the effect of osteopontin (OPN) expression down-regulated by RNA interference (RNAi) on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3.

METHODSHCC-LM3 cells were transfected with the chemically synthesized small interfering RNA (siRNA) formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed.

RESULTSSequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls (P < 0.05).

CONCLUSIONThis study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; Osteopontin ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVEIn this report are reviewed two unrelated patients with typical normokalemic periodic paralysis (normoKPP) features and the results of screening the SCN4A gene for the disease-related mutation.

METHODSTwo sporadic cases with normoKPP were screened for previously known mutations in SCN4A gene (T704M, A1156T, M1360V, I1495F, M1592V) that lead to hyperKPP; denaturing high performance liquid chromatography (DHPLC) was used. Then the rest exons of SCN4A gene were screened by DHPLC, and sequence analysis was performed on those with DHPLC chromatogram variation when compared with unaffected control.

RESULTSTwo cases and one patient's father were detected with V781I, which was proved to be a singular missense mutation in SCN4A gene.

CONCLUSIONThe mutation V781I exists in Chinese patients with normoKPP and may be responsible for normoKPP.

Adult ; Amino Acid Sequence ; Base Sequence ; Child ; Chromatography, High Pressure Liquid ; methods ; DNA ; analysis ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; NAV1.4 Voltage-Gated Sodium Channel ; Paralyses, Familial Periodic ; genetics ; Point Mutation ; Sequence Analysis, DNA ; Sodium Channels ; genetics

OBJECTIVEIn order to explore if power frequency magnetic field (PFMF) can act as cancer promoter or be synergistic with phorbol 12-myristate 13-acetate (TPA) in cancer promotion, the effects of 50 Hz MF on gap junction intercellular communication (GJIC) of astrocytes were observed.

METHODSFluorescence redistribution after photobleaching (FRAP) was adopted to observe the recovery of fluorescence intensity in the bleached cells thus to estimate intercellular communication by gap junction. Comparative fluorescence intensity recovery rate (CFIRR) was as evaluation index. The effects of 50 Hz MF alone or with TPA on GJIC of astrocytes were studied.

RESULTSAfter 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 4.53%/min, whereas that in the control group was 9.74%/min (H = 12.084, P < 0.005). After exposure to 0.8 and 1.6 mT magnetic field for 24 hours respectively, M(d) of CFIRR was 8.25%/min and 6.68%/min respectively, no significant difference from that of control (H = 32.617, P > 0.05). After exposure to 0.8 and 1.6 mT magnetic field for 23 hours then combined with 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 3.32%/min and 2.85%/min respectively, also no significant difference from that in the group treated with 3 ng/ml TPA alone (H = 2.589, P > 0.05).

CONCLUSION50 Hz MF (within 0 - 1.6 mT) alone could not inhibit GJIC of astrocytes; with TPA, could not enhance the inhibition of TPA on GJIC of astrocytes. But with MF intensity increasing, the inhibition of MF on GJIC showed elevated tendency.

Animals ; Astrocytes ; radiation effects ; ultrastructure ; Cell Communication ; radiation effects ; Electromagnetic Fields ; adverse effects ; Gap Junctions ; radiation effects ; Ornithine Decarboxylase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tetradecanoylphorbol Acetate ; pharmacology

OBJECTIVETo investigate the dynamic alterations of neurofilament subunits (NF) in sciatic nerve of hens with organophosphorus ester induced the delayed neurotoxicity or neuropathy (OPIDN).

METHODSHens with OPIDN were produced by giving 30 mg/kg methamidophos subcutaneously to the 10-month-old Roman hens daily for 15 days, and sacrificed after manifesting neurotoxic clinical signs on the 2nd, 10th, and 23rd day respectively. The sciatic nerves were dissected, homogenized and centrifuged. The levels of NF in supernatant and pellet of sciatic nerves were examined by Western blotting respectively at different time from 2 to 23 days.

RESULTSIntegrated optional density (IOD) of high molecular weight neurofilament (NF-H) in sciatic nerve pellet of hens on the day 2, 10, 23 after appearance of OPIDN were 145,117 +/- 17,038, 55,917 +/- 17,333 and 45,038 +/- 6,662 respectively. As compared with the control group (78,875 +/- 22,569), the contents of NF-H in pellet were increased by 84% on day 2, and decreased by 29% and 43% on day 10 and 23 respectively. IOD of NF-H in supernatant of sciatic nerves were 4,709 +/- 1,739, 12,337 +/- 3,205 and 16,745 +/- 931, which were reduced significantly as compared with the control (44,083 +/- 6,895) at three different times. There was no significant difference in IOD of middle molecular weight neurofilament (NF-M) between control group (27,925 +/- 2,660) and on day 2 (31,493 +/- 4,625) in pellet. Those were 19,367 +/- 2,746 and 6,612 +/- 1,119 respectively on day 10 and day 23 in pellet of hen's sciatic nerve, which were much less than that in control. Little were detected in supernatant on day 10, and the IOD of NF-M were 3,196 +/- 269 and 5,206 +/- 1,292 on day 2 and day 23 respectively, which were lessened by 81% and 70% as compared with the control (17,243 +/- 3,232). In sciatic nerve pellet of hens, IOD of low molecular weight neurofilament (NF-L) on day 2 was 39,211 +/- 3,800, which was much higher than that in the control (28,749 +/- 9,319). There were no significant differences between IOD on day 10 (27,974 +/- 3,611), day 23 (21,507 +/- 2,286) and the control. There was no detection both on day 2 and 10 in supernatant of sciatic nerve, and IOD of NF-L were 5,962 +/- 1,929 on day 23, which were reduced significantly compared with the control (11,897 +/- 352).

CONCLUSIONThe alterations of NF in sciatic nerve might contribute to the occurrence and development of OPIDN.

Animals ; Chickens ; Female ; Insecticides ; toxicity ; Neurofilament Proteins ; metabolism ; Organothiophosphorus Compounds ; toxicity ; Sciatic Nerve ; drug effects ; metabolism ; pathology ; Toxicity Tests

OBJECTIVETo study the effects of 1-bromopropane (1-BP) on the functions of learning-memory and the central cholinergic system in rats.

METHODSForty male Wistar rats were randomly divided into four groups: low 1-BP group (200 mg/kg), middle 1-BP group (400 mg/kg), high 1-BP group (800 mg/kg) and control group, and the exposure time was 7 days. The Morris water maze (MWM) test was applied to evaluate the learning-memory function in rats. After the MWM test, the rats were sacrificed, the cerebral cortex and hippocampus were quickly dissected and homogenized in ice bath. The activity of acetylcholine esterase (AChE) and choline acetyltransferase (ChAT) in supernatant of homogenate were detected.

RESULTSThe latency and swim path-length of rats in middle and high 1-BP groups prolonged significantly in place navigation test and the efficiency of searching strategy obviously decreased, as compared with control group (P < 0.05 or P < 0.01). In spatial probe test, the number of crossing platform in three 1-BP groups decreased significantly, as compared with control group (P < 0.05 or P < 0.01). The cortical AChE activity of rats in middle and high 1-BP groups was significantly higher than that of control and low 1-BP group (P < 0.05 or P < 0.01). The AChE activity in rat hippocampus of high 1-BP group obviously increased, as compared with control group as compared with control group (P < 0.05). There was no significant difference of cortical ChAT activity between three 1-BP groups and control group (P > 0.05). In the hippocampus, there was no difference of ChAT activity among the groups (P > 0.05).

CONCLUSION1-BP exposure could significantly influence the learning-memory function in rats due to the increase of AChE activity.

Acetylcholinesterase ; metabolism ; Animals ; Cerebral Cortex ; drug effects ; enzymology ; Choline O-Acetyltransferase ; metabolism ; Hippocampus ; drug effects ; enzymology ; Hydrocarbons, Brominated ; toxicity ; Male ; Maze Learning ; drug effects ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of tacrolimus (FK506) on behavioral function and heat-shock proteins (HSP70) expression in nervous tissues of acrylamide (ACR)-induced rats.

METHODSTotally 40 health Wistar rats were randomly divided into control group, model group, low and high doses of FK506 groups. All four groups were treated five times per week for four weeks. Gait score was measured every week. And rats were sacrificed on day 28, the cerebrum, spinal cord and sciatic nerve were dissected, and homogenized in ice bath, then the levels of HSP70 and Bcl-2, Bax were analyzed by western bloting.

RESULTSCompared with the ACR model group, the gait score in low and high doses of FK506 groups decreased by 30.1% and 47.7% respectively in the 4th week. In the cerebrum and sciatic nerve pellet, the level of HSP70 in the FK506 groups increased by 11.6%, 33.3% and 56.3%, 58.5% (P < 0.01), but no significant changes existed in spinal cord. The level of Bcl-2 in the sciatic nerve pellet increased by 39.1% (P < 0.01) but no significant changes existed in the cerebrum and spinal cord from low dose of FK506 group. And the level of Bax in the spinal cord pellet markedly increased by 46.8% but not in cerebrum and sciatic nerve pellet; Whereas in the tissues mentioned above, the levels of Bcl-2 were enhanced remarkably by 16.3%, 14.8% and 56.0% (P < 0.01) in the high dose of FK506 group. And the level of Bax in the cerebrum and spinal cord pellet markedly increased by 16.4% and 40.2% but not in sciatic nerve. The values of Bcl-2/Bax in low and high doses of FK506 groups clearly increased by 15.9%, 33.3%, 36.9% and 30.1%, 49.1%, 60.1% (P < 0.01).

CONCLUSIONThe administration of FK506 has dramatically neuroprotective effects against the development of ACR neuropathy, which may be related to up-regulating the expression of HSP70 and Bcl-2 with down-regulating the expression of Bax.

Acrylamide ; poisoning ; Animals ; HSP70 Heat-Shock Proteins ; metabolism ; Male ; Nerve Tissue ; drug effects ; metabolism ; Neuroprotective Agents ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Tacrolimus ; therapeutic use ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo compare the long-term results of procedure for prolapse and hemorrhoids(PPH) and Milligan-Morgan(MM) hemorrhoidectomy in the treatment of third- and fourth-degree hemorrhoids.

METHODSNinety-one patients in PPH group and 120 patients in MM group, treated in our hospital from May 2001 to May 2005, were followed up. All the patients suffered with third- or fourth-degree hemorrhoids. The data including symptoms relief, complication and recurrence were compared between two groups.

RESULTSPPH and MM both significantly relieved bleeding (95.6% vs 92.7%), pain (93.1% vs 94.3%) and prolapse (93.4% vs 93.1%). There were no significant differences between two groups(P >0.05). PPH significantly relieved constipation compared with MM(60.0% vs 32.1%, P <0.05). No significant differences of complete relief rates between PPH group and MM group according to different degrees, types and prolapses of hemorrhoids were found(P >0.05). There were no significant differences in the overall complication rate(14.3% vs 12.7%), recurrence rates(12.1% vs 8.8%) and satisfactory degree (85.1 vs 88.2) between two groups as well.

CONCLUSIONThe long-term outcomes of PPH in the treatment of third and fourth-degree hemorrhoids are similar to Milligan-Morgan hemorrhoidectomy.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Digestive System Surgical Procedures ; methods ; Female ; Follow-Up Studies ; Hemorrhoids ; surgery ; Humans ; Male ; Middle Aged ; Surgical Stapling ; Surgical Stomas ; Young Adult