Objective To study the relationship between the intervals of Mitomycin C treatment and cytotoxicity, apoptosis and drug resistance for bladder cancer cells.Methods The bladder transitional cell cancer line BIU-87 was treated for two hours every time for five times with intervals of 24, 48, 72 and 96 hours respectively.Cytotoxicity was measured by MTT.p53,bcl-2,Bax and p170 expression were analyzed by Western blot.Results The IC_(50)(?g/ml)were 4.41,0.71,2.83,4.51and 6.16 with treatment intervals of 24, 48, 72 and 96 hours respectively, p53 and bcl-2 were significantly down-regulated and bcl-2/Bax was re- duced at 24 hour treatment interval but not changed at 48,72 and 96 hour intervals,p170 was not detected at 24 hour treatment interval but increasingly expressed at 48,72 and 96 hours intervals.Conclusion The in- terval of Mitomycin C treatment is closely related with cytotoxieity and apoptosis and drug resistance of blad- der cancer cells.The intervals of intravesical instillations may play an important role in the effect of chemotherapy.

OBJECTIVETo investigate the mechanism of lemon peel essential oil (LPE) on the cariogenicity of Streptococcus sobrinus (Ss).

METHODSLPE was extracted by the authors, and the minimum inhibition concentration (MIC) was measured by disc diffusion method. The LPE was used as the experimental group with concentrations ranging from 2.250 g/L to 0.281 g/L prepared with trypticase peptone yeast (TPY) culture medium, and TPY culture medium was used as the control group. Ss at the concentration of 10(8) CFU/ml was added to each group, and cultured for 6, 18, 24, 48 hours. Neson-Somogyi method was used to measure the content of reducing sugar, and glucosyltransferase (GTF) activity. The activity of lactate dehydrogenase (LDH) was measured by lactic acid and pyruvic acid continuous monitoring method. The content of water insoluble glucan (WIG) was measured by anthrone method, and the pH value of the culture solution was detected. The value of pH before the experiment and the time difference was alculated as ΔpH.

RESULTSAt the same time point, the activity of GTF and LDH and the concentration of WIG and the value ΔpH decreased gradually with the increase of concentration of LPE. There were significant differences between each experimental group and control group (P < 0.01). The control group had the maximum value, GTF: (6.71 ± 0.61) mIU, LDH: (135.8 ± 1.7) U/L, WIG: (47.15 ± 5.12) mg/L, ΔpH: (2.67 ± 0.01). The highest drug concentration group had the minimum value: GTF: (0.39 ± 0.07) mIU, LDH: (95.0 ± 5.4) U/L, WIG: (2.44 ± 0.38) mg/L, ΔpH: (0.61 ± 0.01).

CONCLUSIONSThe LPE below the MIC could still inhibit the GTF, LDH activity and lead to the decrease of WIG and the acid production.

Dose-Response Relationship, Drug ; Glucans ; biosynthesis ; Glucosyltransferases ; antagonists & inhibitors ; metabolism ; Lactate Dehydrogenases ; antagonists & inhibitors ; metabolism ; Microbial Sensitivity Tests ; Oils, Volatile ; pharmacology ; Plant Oils ; pharmacology ; Streptococcus sobrinus ; drug effects ; metabolism

This study was aimed to investigate the chimerism and fusion gene expression in patients with CML after allo-HSCT, to analyse engraftment and minimal residual disease by using STR-PCR combined with RT-PCR qualitative and quantitative assays, and to evaluate their clinical value for predicting disease relapse. 4 relapsed patients with CML after allo-HSCT were dynamically investigated. Qualitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was performed by RT-PCR. The results showed that the 100% donor chimerism appeared in 4 patients on day 28 after transplantation and bcr/abl expression was negative, but the 4 patients were in status of unstable mixed chimerism (DC: 0% - 80.4%) at the different time points during the following up with bcr/abl gene positive. 2 patients of them were continuously mixed chimerism after relapse of CML, the other 2 changed from MC to CC by intervention of clinical treatment. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and CML relapse, and bcr/abl gene expression was positive. It is concluded that the results of STR-PCR in the range of its sensitivity fully correspond with bcr/abl tests in patients. The combination of STR-PCR with RT-PCR will provide a highly sensitive and valuable tool for evaluating engraftment, graft rejection, and relapse and predicting GVHD. Furthermore, it can provide a basis for early intervention of clinical treatment, and can identify these high risk patients with molecular or cytogenetic relapse after allo-HSCT.
Fusion Proteins, bcr-abl ; genetics ; metabolism ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Neoplasm Recurrence, Local ; genetics ; Neoplasm, Residual ; diagnosis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Transplantation Chimera ; Transplantation, Homologous

OBJECTIVETo study the effects of Erlong Zuoci Pill (, ELZCP) and its disassembled: prescriptions on gentamicin (GM)-induced ototoxicity model in vitro.

METHODSAfter the spiral organ of cochleae: of newborn mice (postnatal days: 2-3) cultured for 24 h, GM alone or combined with water extracting-alcohol precipitating solution of ELZCP or with its disassembled prescriptions was added. Hair cells were observed under a fluorescence microscope after TRITC-phalloidin staining, and the cochlear hair cell loss rate was calculated by counting the whole cochlear hair cells and analyzed by whole cochlear hair cells analyzing software.

RESULTSGM induced cochlear outer hair cells (OHCs) and inner hair cells (IHCs) injuries in a dose-dependent manner, and they were significantly different as compared with those in the normal control group (P<0.05, P<0.01). ELZCP at the concentration of 0.003-3 mg/mL could decrease the hair cells loss induced by the 0.3 mmol/L GM (P<0.05, P<0.01), the effects was in a dose-dependent manner, and the concentration of 0.3 mg/mL showed the optimal protective effect. For the ELZCP disassembled prescriptions, Liuwei-Dihuang could decrease OHC loss rate than that in the 0.3 mmol/L GM model group (P<0.05), but the OHC loss rate was still higher than that in the ELZCP group (P<0.01), which indicated that the protective effect of hair cells by Liuwei-Dihuang was not better than that of ELZCP. Poria decreased OHC loss rate from 72.1 % +/-3.7 % to 58.8 %+/- 8.2 % (P<0.05).

CONCLUSIONSELZCP could play a role in antagonizing the injury of cochlear hair cells induced by GM ototoxicity,: and its disassembled prescriptions, Liuwei-Dihuang was the main component to protect the cochlear hair cells from GM-induced ototoxicity, and Magnetitum combined with Radix Bupleurui could strengthen the action of the whole prescription; Poria could reduce GM-induced OHC loss.

Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Gentamicins ; toxicity ; Hair Cells, Auditory, Inner ; drug effects ; pathology ; Hair Cells, Auditory, Outer ; drug effects ; pathology ; Mice ; Organ of Corti ; drug effects ; pathology ; Prescriptions ; Tablets

OBJECTIVETo discuss the causes, sites, management strategies and curative effects of accidental facial nerve paralysis in the middle ear surgery.

METHODSForty two cases with peripheral facial nerve paralysis following middle ear surgery who underwent surgical exploration and reanimation were analyzed. Facial nerve decompression, primary end-to-end anastomosis, interpositional nerve grafts with the great auricular nerve and nerve substitution of facial-hypoglossal anastomosis were applied to restoration of the facial nerve function. The facial nerve function was graded according to House-Brackmann (HB) Grade.

RESULTSThe most common operation complicating iatrogenic facial nerve injury was mastoidectomy, and the common sites of the injured facial nerve were the tympanic segment and pyramid segment. The facial nerve exploration showed facial nerve edema in nine cases (21.4%), injury of the facial nerve sheath was observed in 10 cases (23.8%), partial nerve fibers transection was found in four cases (9.5%), total nerve fibers transection was detected in 17 cases (40.5%) and two cases (4.8%) with facial nerve anatomical integrity. Facial nerve re-animation methods include facial nerve decompression in 24 cases (57.1%), end-to-end anastomosis in two cases (4.8%), end-to-end anastomosis after nerve transfer in two cases (4.8%), interpositional nerve grafts with the great auricular nerve in 10 cases (23.8%) and facial-hypoglossal nerve anastomosis in four cases (9.5%). The facial nerve function was graded according to House-Brackmann Grade before and after surgery. Twenty eight patients were followed up more than one year. For the 17 cases who received facial nerve decompression, four cases recovered to House-Brackmann Grade I, 11 cases recovered to House-Brackmann Grade II, two cases recovered to House-Brackmann Grade III. For the five cases who underwent the great auricular nerve grafting, three cases recovered to House-Brackmann Grade II, two cases recovered to House-Brackmann Grade III. For the four cases who received facial-hypoglossal nerve anastomosis recovered to House-Brackmann Grade III. For the two cases who underwent the end-to-end anastomosis recovered to House-Brackmann Grade II.

CONCLUSIONSThe tympanic segment and pyramid segment are more vulnerable to be injured during mastoid surgery. The injured facial nerve should be explored and repaired. The methods include facial nerve decompression, end-to-end anastomosis, end-to-end anastomosis after nerve transfer, interpositional nerve grafts with the great auricular nerve and facial-hypoglossal nerve anastomosis.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Ear ; surgery ; Ear, Middle ; surgery ; Facial Nerve Injuries ; diagnosis ; etiology ; surgery ; Female ; Humans ; Iatrogenic Disease ; Intraoperative Complications ; Male ; Mastoid ; surgery ; Middle Aged ; Otologic Surgical Procedures ; adverse effects ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the risk factors of road injury.

METHODSCase-control study was used. From November 2001 to August 2002, 406 drivers who had 438 drivers who had not experienced a motor vehicle crash in Huanggu district, Shenyang city were recruited by randomly selection on time of day, day of week and site in the same period at same district. Face to face interviews with drivers were conducted according to a highly structured questionnaire covering the circumstances of the current trip, usual behavior and background characteristics of the drivers and the condition of motor vehicles. Stanford sleepiness scale and Epworth sleepiness scale were used to quantify acute and chronic sleepiness respectively.

RESULTSIncreased risk was associated with drivers who identified themselves as having chronic doziness (OR = 1.98, 95% CI: 1.26 - 3.12). Increase in risk was associated with measures of acute tiredness, but without statistical significance (OR = 2.38, 95% CI: 0.89 - 6.31). Comparing to permanent daytime work pattern, rotating shifts or permanent night-work pattern increased the risk of crash (OR = 2.09, 95% CI: 1.48 - 2.94). The risk of motor vehicle crash among the drivers who drank alcohol in the previous 6 hours was 3.59 times (95% CI: 1.13 - 11.39) of those drivers who did not drink. Driving violations also contributed to the increased risk of crash (OR = 1.73, 95% CI: 1.22 - 2.46).

CONCLUSIONFactors as chronic doziness, rotating shifts or permanent night-work pattern, driving under alcohol impairment, violation of motor vehicle regulation all significantly increased the risk of road injury. Acute sleepiness might serve as a potential risk factor for road injury.

Accidents, Traffic ; Adult ; Automobile Driving ; Case-Control Studies ; Female ; Humans ; Logistic Models ; Male ; Risk Factors

This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.
Animals ; BCG Vaccine ; immunology ; CHO Cells ; Cricetinae ; Cricetulus ; Granulocyte-Macrophage Colony-Stimulating Factor ; immunology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Rats ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo study the genetic stability of an immortalized cell line transformed by Epstein-Barr virus (EBV) after long subculture process.

METHODSIn the present study, the genetic stability including chromosome diploidy, karyotypes and microsatellite DNA were evaluated with chromosome banding techniques and microsatellite DNA detection. The telomerase activity of the immortalized cell line was detected by using the telomerase assay kit.

RESULTSFrom passage 1 to 30, there were no change of the diploidy, karyotypes of chromosome and microsatellite DNA, and the telomerase activity is negative.

CONCLUSIONThis study indicates that the immortalized cell line remains stable genetically within limited passages.

Cell Transformation, Viral ; genetics ; Herpesvirus 4, Human ; genetics ; Humans ; Lymphocytes ; cytology ; metabolism ; virology ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction

OBJECTIVETo generate an oncolytic herpes simplex virus (oHSV) permissive mouse melanoma cell line B16RHSV, preserving the tumorigenic ability in syngeneic mice.

METHODSThe herpes simplex virus entry mediator (HVEM) gene was amplified by PCR from human melanoma cell line A375, and cloned into pGEM-T Easy vector for sequencing. The HVEM gene was then cloned into pcDNA3 vector to generate pcDNA3-HVEM for transfection of mouse melanoma cell line B16-F10 cells. After that, the putative transfected cells were selected in full growth medium containing G418. The HVEM-expressing cells were isolated by immunomagnetic bead separation. The mouse melanoma cell line expressing oHSV receptor-HVEM, designated as B16RHSV, was generated. The permissibility of B16RHSV cells to oHSV infection was examined with green fluorescence protein (GFP)-expressing oHSV (oHSVGFP). To investigate the tumorigenic ability of both cells in vivo, 2×10(5) cells in 100 µl were subcutaneously inoculated into the right flanks of C57/BL mice.

RESULTSIn vitro, the B16RHSV mouse melanoma cells were shown by fluorescence microscopy capable of being infected by oHSVGFP. In vivo, the B16RHSV cells, like their wild type counterpart, grew to form melanoma in syngeneic mice.

CONCLUSIONA herpes simplex virus-permissive mouse melanoma cell line was established. Its tumorigenicity remained unchanged.

Animals ; Cell Line, Tumor ; Female ; Gene Amplification ; Genetic Vectors ; Herpesvirus 1, Human ; genetics ; physiology ; Humans ; Melanoma ; pathology ; virology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Plasmids ; Receptors, Tumor Necrosis Factor, Member 14 ; genetics ; metabolism ; Transfection ; Tumor Burden