Objective To study the effects of chimeric B7 antibodies on mononuclear cells from patients with ankylosing spondylitis (AS).Methods Chimeric antibodies of ch4E5 and chlD1 against B7 with a final concentration of 5 μg/ml were respectively added into culture media of mononuclear cells from patients with AS.Two other groups including huIgG-treated and untreated culture medium of mononuclear cells from patients with AS were set up as the controls.The culture medium of mononuclear cells from healthy subjects were prepared as normal control group.The disparities of mononuclear cell death in culture among each group were analyzed by using micro-lymphocyte cytotoxicity test (MLCT).The expression of membrane molecules were analyzed by FCM.The concentrations of cytokines in culture media were measured by ELISA.Results Both groups treated with the chimeric antibody showed lower scores for cell death than untreated group as indicated by MLCT (P＜0.01).FCM analysis demonstrated that both CD4/CD8 ratio (P ＜0.05) and membrane expression of CD154 (P＜0.01) decreased in chimeric antibody treated groups,but the number of CD4+CD25high Treg cells (P＜0.01) increased as compared with those in untreated group.The concentrations of IL-2 and TNF-α were down-regulated (P＜0.05) in chimeric antibody treated groups,while IL-4 and TGF-β were up-regulated (P＜0.02) as compared with those in untreated group.Conclusion Chimeric B7 antibodies might affect the expression of T cell membrane molecules and the cytokine production by mononuclear cells through B7/CD28 and CD4O/CD154 signaling pathways in patients with AS.

Objective To explore a novel surgical treatment for chronic suppurative otitis media (CSOM) and evaluate its treatment effect.Methods All 97 patients with chronic suppurative otitis media were chosen to be treated using this new surgical method.The skin of the external auditory canal was maintained intact.Open radical mastoidectomy was used to complete clean-up lesions ; the fascia of pedicled temporalis myofascia (PTM) was used to repair the tympanic membrane.The pedicled temporalis fascia,pedicled postauricular periosteal flap and intact skin of the external auditory canal were used in reconstruction of the posterior wall of external auditory canal.Pure tone audiometry (PTA) was performed before and after surgery,recording the air conduction and bone conduction thresholds at 0.5 kHz,1.0 kHz,2.0 kHz,4.0 kHz.The average of the patient's air and bone conduction hearing thresholds was recorded at the 4 frequencies.External auditory canal gauze was removed 3 weeks after surgery.All subjects were followed up for over 2 years.Comparison of hearing thresholds (PTA) was made ① Before and 4 weeks after surgery.② Before and 2 years after surgery.Hearing function comparison include air conduction (AC),bone conduction (BC) and air-bone gap (ABG) analysis.SPSS 16.0 was used in statistical analysis.Pre-and postoperated AC,BC and ABG were compared using T-test.P ＜ 0.05 was considered statistically significant.Results The healing rate of post-operated tympanic membrane was 95.88％ (93/97).Ninty-six ears had 2-year follow-up,and 1 patient was lost in follow-up.There were 2 patients presented with eardrum perforation during the follow-up,and the 2-year healing rate of tympanic membrane perforation was also 93.85％ (92/97).In 96 ears with 2-year followed-up,the average of pre-AC was (52.10 ±3.96) dB,the average of post-AC was (35.67 ±2.52) dB; the average of preABG was (36.6 ± 5.2) dB,and the average of post-ABG (± SD) was (12.14 ± 6.20) dB.Statistical analysis showed significant difference between preoperative and postoperative AC or ABG values (P ＜ 0.05).Conclusion The present surgical procedure broke through the existing conventional mastoidectomy of making a surgical incision in the posterior wall of the external auditory canal.This procedure cleared the lesion completely and preserved the physiological function of the external auditory canal.The acoustic systems and state of gasification to the mastoid tymnpanum were reconstructed,rehabilitated and maintained.The healing rate of hearing and tympanic membrane perforation was improved.

OBJECTIVE: To investigate the optimizing conditions in isolation of the side population in laryngeal carcinoma cell line Hep-2. METHOD: Single-cell suspension cells were detached from the culture flask with trypsin EDTA, at a concentration of 1 x 10(6) cells/ml. (1) The trail Samples were incubated with Hoechst33342 at a concentration of 5 microg/ml, 9 microg/ml, 10 microg/ml, 11 microg/ml for 90 minutes. (2) They were incubated with Hoechst for 50, 70, 90, 110, 130 min in water bath individually. (3) The single-cell suspension were incubated Hoechst in water bath and in thermostat each. (4) The two different density of cells were harvested, which were 100% and 70%, and then di gest into single-cell suspension. Once incubation finished, suspended in phosphate buffered saline (PBS), then test SP% by flow cytometry. Among all groups,Verapamil hydrochloride was added to the control samples, incubated at 37 degrees C for 30 minutes, the other condition were keep the same with their trial groups. RESULT: (1) The percentage of Hoechst-negative cells in trial group was (39.96 +/- 0.24)%, (26.23 +/- 0.39)%. (18.79 +/- 0.02)%, (19.01 +/- 0.14)% at the concentration of 5 microg/ml, 9 microg/ml, 10 microg/ml, 11 microg/ml respectively, when the PI-positive cells were (30.45 +/- 0.63)%, (49.9 +/- 0.42)%, (50.12 +/- 0.68)%, (64.16 +/- 0.39)% separately. (2) Varying the duration of staining incubation showed that there was a typical FACS pattern and SP% was constant when the incubation was at least 90 min. (3) Compare to water bath, SP% was more than in thermostat, the SP% was (18.67 +/- 0.45)%, (22.6 +/- 0.50)% respectively; (4) Cell density is also responsible for SP%. The low density the cell is, the less in SP%. SPSS13.0 was used in statistical analysis, the groups were compared using t-Test. P < 0.05 was considered statistically significant. CONCLUSION The optimum concentration and duration of incubation of Hoechst33342 in isolation of the side population cells in laryngeal carcinoma cell line Hep-2 is 10 microg/ml and 90 min. Incubated in water bath is better than in thermostat. The best staining cell density is around 80%-90%.
Cell Count ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; pathology ; Neoplastic Stem Cells ; cytology ; Side-Population Cells ; cytology

OBJECTIVE: To explore the possibility mechanism of non-side population cells (NSP) of Hep-2 be induced into stem-like cancer cells by chemotherapy drug--cisplatin. METHOD: Hep-2 cell lines were sorted by fluorescence-actived cell sorting. The acquired NSP cells in trail group were co-cultured with cisplatin for more than 48 hours,while the control group with normal saline(NS). Then identified the percentage of the side population (SP) cells by flow cytometer. The β-catenin, notch-1 mRNA in trial and control group were detected using quantitative realtime PCR, and the β-catenin, notch-1 protein in two groups were compared by Western blot. RESULT: The percentage of side population cells in two groups were (17.16 ± 0.18)%, (10.05 ± 1.20)%, respectively. There was significant difference between two groups (t = 5.844, P < 0.01). The expression of β-catenin, notch-1 was higher in trail group by qRT-PCR; the protein levels of β- catenin, notch-1 was found to inceased in the trail group by Western blot (t = 5.155, P = 0.031; t = 5.977, P = 0.004). Statistical analysis showed significant difference between two groups (P < 0.05). CONCLUSION NSP cells can be differentiated into stem-like cancer cells after being treating with cisplatin. The supposed mechanism is maybe through wnt/β-catenin, notch signaling transduction pathway abnormalities.
Antineoplastic Agents ; pharmacology ; Cell Differentiation ; Cell Line, Tumor ; Cell Separation ; Cisplatin ; pharmacology ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; pathology ; Neoplastic Stem Cells ; drug effects ; RNA, Messenger ; Signal Transduction ; beta Catenin

OBJECTIVE: To investigate the changes of laryngeal cancer Hep-2 cells to cisplatin chemosensitivity after the interference of siRNA of β-catenin gene expression. METHOD: Using a small interference RNA (siRNA) technology interfere β-catenin gene of Hep-2 cells . The mRNA and protein levels of β-catenin in the Hep-2 cells of different groups were detected by qPCR and Western blot. It was divided into siRNA-β-catenin-Hep-2 siRNA group, β-catenin-Neg negative control group and blank control group. Cell proliferation inhibition rate of different concentrations of cisplatin on three groups was detected by MTT assay. Calculate the 50% inhibitory effective concentration IC50 value. Check the change of three groups of cells' apoptosis rate by flow cytometry after the same concentrations of cisplatin stimulation. RESULT: β-catenin-siRNA interference fragment can specifically reduce the expression levels of β-catenin mRNA and protein. qPCR illustrated the expression of mRNA in β-catenin-siR-NA-Hep-2 interference group decreased 70% (P < 0.05) compared with the control group, Western blot results showed that the β-catenin protein expression of interference group (0. 545 ± 0.111) decreased significantly compared with blank control group (1.507 ± 0.139) and negative control group (1.429 ± 0.089), P < 0.05. The IC50 calculation software showed that IC50 of cisplatin on β-catenin-siRNA IC50 interference group is (5.81 ± 0.46)μg/ml, the blank control group is (10.10 ± 1.01) μg/ml, the difference between the two groups has statistical signifi- cance (P < 0.01). Cell apoptosis rate of β-catenin-siRNA interference group was (26.15 ± 0.60)%, significantly higher than the control group (14.16 ± 0.05)%, P < 0.05. CONCLUSION To interfere the expression of β-catenin can effectively enhance the sensitivity of laryngeal cancer cells to chemotherapeutic drugs cisplatin. It provides a theoretical support for the reduction of laryngeal cancer chemotherapy drug cisplatin dosage.
Apoptosis ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Humans ; Laryngeal Neoplasms ; genetics ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; beta Catenin ; genetics

OBJECTIVE:To provide reference for medical costs control in China. METHODS:Based on main characteristics of the United States health service system,information asymmetry,principal-agent theory and Freedman’s consumption theory were used to analyze the advantages of restrictive relationship among doctors,hospitals and health insurance institutions in control-ling medical costs and improving the quality of diagnosis and treatment. The growth rate of medical cost,the percentage of drug ex-penditure and other aspects were compared between China and the United States;the effect of restrictive relationship on medical cost control was demonstrated. RESULTS & CONCLUSIONS:In the United States,there are hierarchical medical system and two-way referral system;for-profit hospitals and non-profit hospitals are mutually complementary;different natures of health insur-ance system,different payment methods and strict“commercial bribe”monitoring system are carried out;doctors,hospitals and health insurance restrict each other. Not only there are many advantages in theory,but also in practice the growth rate of medical cost and the percentage of drug expenditure are superior to our country. Finally it controls the rapid growth of medical cost to a cer-tain extent. Combined with our national conditions,learning from the United States experience,restrictive relationship among doc-tors,hospitals and health insurance institutions is established to control the increase of medical cost in China through reducing infor-mation asymmetry and standardizing payment audit;establishing a scientific pattern of mixed payment;strengthening the indirect impact of the health insurance institutions on doctors and hospitals,etc.

Objective To explore the related pelvic floor anatomy to the pathological vaginal relaxation and key points of the vaginal tightening surgery.Methods The vaginal tightening surgery was performed in 24 cases of vaginal relaxation.The key points of this operation included levator ani muscle suturation and perineal body reconstruction,and anal sphincter reconstruction as in case of the muscle injury grade Ⅲ.Results The degree of levator ani muscle separation was positively correlated with that of vaginal relaxation in all the 24 cases.18 cases were followed up from 6 months to 2 years,and had no complications of rectovaginal fistula and infections.The average level of perineal body was increased from 2.3 cm to 3.5 cm.Vaginal length of 6 cm from vaginal orifice was proper with good tightness.The patients felt strong anal contraction,enhanced ability of vaginal tightening and improved quality of sex life.There were no more infections of genitourinary tract.Conclusions Through levator ani muscle suturation and perineal body reconstruction,it can get the vaginal tightening effect.

The infection caused by pseudomonas aeruginosa biofilm has becoming one of the most urgent problems in hospital.Recent reports of possible mechanism of iron in biofilm formation were reviewed in this paper.The possible effects of iron on adsorption,microcolony formation,mature of colony and desorb were explained,and the prospect of clinical use of local iron or chelator spray was also reviewed here.

To employ multiplex polymerase chain reaction (PCR) to classify the types of virus in 128 patients of community-acquired pneumonia and analyze the relationship between type of virus,age and seasons.The positive rate of viral pneumonia was 34.4％ (44/128).In 44 virus positive patients,the rates of influenza A virus,rhinovirus,adenovirus,parainfluenza type 1,human metapneumovirus virus,influenza B virus,parainfluenza type 2,parainfluenza type 3 and enterovirus were 22.7％ (10/44),15.9％ (7/44),15.9％ (7/44),9.1％ (4/44),9.1％ (4/44)6.8％ (3/44),2.3％ (1/44),2.3％ (1/44) and 2.3％ (1/44) respectively.And other 6(13.6％) patients were at least infected by one virus.The viral pneumonia had a high positive rate in spring and winter.And the age group of 20 to 29 years had a high positive rate.The major pathogen contributing to adult community-acquired pneumonia in Yantai were influenza A virus,rhinovirus and adenovirus.

Objective To study the relationship between rs290487,rs7903146 of transcription factor 7-Like 2 (TCF7L2) gene and post-transplantation diabetes mellitus in Han Zhejiang population.Method We genotyped two single nucleotide polymorphisms (SNPs) across the TCF7L2 gene in 90 unrelated post-transplantation type 2 diabetes mellitus (PTDM) patients,112 unrelated non-PTDM patients,and a set of post-transplantation diabetes mellitus patients (n =68).Genotyping was performed using direct sequencing SNP Genotyping Assays.The association of SNPs with post-transplantation diabetes mellitus was analyzed.Result In this study,there was statistically significant difference in the T-allele of TCF7L2.rs7903146 between PTDM group (5.1％) and non-PTDM group (1.3％) (P＜0.05).For rs7903146,the frequencies of genotype C/C,C/T (70.0％) and T/T (35.8％) was statistically significant in PTDM group (P＜0.05).For rs290487,the frequencies of genotype C/C,C/T and T/T was 14.7％,38.2％ and 47.1％ respectively in PTDM group,P＞ 0.05.The incidence of PTDM was significantly higher in patients with the CT genotype (odds ratio 18.54 [95％ CI 1.21-282.26],P =0.03).Conclusion With the current sample size,we found that the CT genotype of rs7903146 was significantly associated with post-transplantation diabetes mellitus.