Adult ; Aged ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Knee Injuries ; surgery ; Male ; Middle Aged ; Postoperative Care

BACKGROUND:intervertebral disc degeneration can reduce nucleus pulposus cells,and peroxiredoxin II involved in the regulation of resist oxidation damage,cell division,differentiation,signal transduction and apoptosis.Peroxiredoxin Ⅱ has promotive effect on intervertebral disc degeneration,whereas the mechanism remains poorly understood.OBJECTIVE:To observe the effects of perexiredoxin Ⅱ on human intevertebral disc cells activity and type Ⅱ collagen synthesis in vitro.METHODS:Human degenerated human lumbar disc cells were cultured in vitro,and assigned into the control and peroxidase Ⅱ groups.Peroxidase Ⅱ with doses of 10,100 and 1 000 ng/L were added into the peroxidase Ⅱ groups.The cells were identified by immunohistochemical staining,and the cell proliferation was detected using cck-8 kit.Cell supematant was collected at days 3 and 7 after operation,and the expression of type Ⅱ collagen was measured by double-antibody sandwich enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:In vitro cultured human degenerative lumbar intervertebral disc nucleus pulposus cells by adding peroxidase increased with the dose-Ⅱ,the disc nucleus pulposus cells of the volume and type Ⅱ collagen synthesis gradually reduced(P ＜ 0.01).Tips peroxidase Ⅱ on the intervertebral disc nucleus pulposus cells,the number and type Ⅱ collagen synthesis significantly inhibited in a dose-dependent manner.Thus speculated that peroxidase Ⅱ on the nucleus pulposus cells in vitro may lead to disc degeneration as a precipitating factor.

Objective To investigate the expressions of CK17 and P63 in cervical cancer tissues,and explore the possibility of CK17 and P63 as markers of cervical cancer stem cells.Methods The expressions of CK17 and P63 in cervical squamous cancer tissues(n=55) and normal cervix tissues(n=20) were detected with immunohistochemical method.Results ①CK17 expressed in reserve cell cytoplasm in normal cervical specimens.In the cervical cancer specimens,the expression of CK17 was increased and it seemed that CK17 was stained in the margin of the cancer nest and the tumor embolus.In specimens with high grade malignant cancer or after the chemotherapy,the expressions of CK17 were increased(P

Objective To explore the relationship between peripheral Th17 cell number and chronic gastritis in Helicobacter pylori(H.pylori)-infected children.Methods Children were diagnosed as chronic gastritis by endoscopy.The degree and activity of inflammation were graded by histopathology examinations.The patients with both 13C urea breath test and urease test positive were diagnosed as H.pylori infection.The peripheral Th17 cell number was measured by flow cytometry and expressed as a ratio to total T cell.Results The Th17 cell number in HP group (chronic gastritis with H.pylori infection,n =33),non-HP group (chronic gastritis without H.pylori infection,n =24) and normal controls (n =15) were (1.55 ±0.30)％,(1.06 ±0.33)％,and (1.04 ±0.35)％,respectively.HP group included a statistically higher Th17 cell number than the other groups (all P ＜ 0.05),while no obvious difference was found between non-HP group and controls (P ＞ 0.05).According to the degree of inflammation,the chronic gastritis with H.pylori infection was categorized into non-apparent (n =10),mild (n =8),moderate (n =9) and severe (n =6) subgroups.The Th17 cell number in each subgroup was (1.64 ± 0.21)％ (non-apparent),(1.61 ± 0.23)％(mild),(1.25 ± 0.29) ％ (moderate) and (1.75 ± 0.20) ％ (severe),respectively.The moderate group had a lowest Th17 cell number among 4 groups (P ＜ 0.05).And significant differences did not exit in the other 3 groups (P ＞ 0.05).The HP group patients with different inflammatory activity had a Th17 cell number of (1.23 ±0.25)％ in nonapparent (n=15),(1.53 ±0.15)％ in mild (n=6),(1.55 ±0.32)％ in moderate (n=6) and (1.71 ±0.35)％ in severe (n =6) subgroup,respectively.However,there were no significant differences among 4 subgroups (P ＞ 0.05).Conclusions In the progress of chronic gastritis with H.pylori infection,Th17 cells may play a role as a double-edged sword by protecting and fighting against H.pylori infection and immunopathologic insults.This would provide more insights into the treatment of H.pylori infection.

Objective To explore the application of reading education in cultivating nursing students' humanities in vocational college.Methods Nursing students were selected at random and divided into two groups,35 students in the experimental group and 50 in the control group.The students in the experimental group were given the 2-year related instruction through reading activities in the light of the principle enhance general humanities foundation in the first grade and strengthen humanistic features in the second grade.Nursing skill assessment and questionnaire oriented to humanities were used to test the educational effect.Results The result of nursing skill assessment in the experimental group was significantly better than that of the control group.The score of humanities' moral,law,culture and aesthetic dimension and the total score of the experimental group were significantly better than those of the control group.Conclusions Well targeted reading is effective in the cultivation of nursing students' humanities.Meanwhile reading education can serve as an important and beneficial way to improve the cultivation mode of vocational college's nursing humanities.

Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.

Objective To explore the changes of programmed death receptor-1 (PD-1) in chronic hepatitis B (CHB) patients with different baseline of hepatitis B virus (HBV) DNA levels after treatment with adefovir dipivoxil (ADV).Methods One hundred CHB patients with positive hepatitis B e antigen (HBeAg),1 × 104 copy/mL≤HBV DNA≤1 × 107 copy/mL,and positive human leukocyte antigen-A2 were divided into two groups according to the baseline HBV DNA level:47 cases in low virus load group whose HBV DNA level was ≤1 × 105 copy/mL; 53 cases in high virus load group whose HBV DNA level was＞1 × 105 copy/mL.Both groups were treated with ADV 10 mg/d.Serum HBV DNA,HBeAg seroconversion rate,alanine aminotransferase (ALT) and total bilirubin (TBil) levels of both groups before treatment and 12 months after treatment were compared.Flow cytometry was used to test peripheral blood HBV-specific cytotoxic T lymphocyte (CTL) surface PD-1 and peripheral blood HBV-specific CTL level.Categorical data were tested by x2 test; quantitative data was compared with t-test.Results Peripheral blood HBV-specific CTL surface PD-1 of CHB patients in low virus load group was 20.17 ％±1.69％,which was lower than that in high virus load group (41.38％±2.30％,t =53.02,P＜0.01) ; peripheral blood HBV specific CTL levels in two groups were 0.37％±0.02％ and 0.17％± 0.02％,respectively (t=50.47,P＜0.01) ; ALT and TBil levels in low virus load group were both lower than those of high virus load group (t=13.07,P＜0.01; t=5.06,P＜0.01).Twelve months after treatment,HBV DNA of 25 cases (53.2％) in low virus load group and 10 cases (18.9％) in high virus load group were lower than the detectable level (HBV DNA＜500 copy/mL,x2 =12.89,P＜0.01);HBeAg seroconversion was achieved in 15 cases(31.9％) and 1 case (1.9％),respectively (x2 =16.72,P＜0.01) ; peripheral blood HBV-specific CTL surface PD-1 expression levels were 9.00 ％ ±1.38 ％ and 29.40 ％ ± 3.76 ％,respectively (t =36.80,P＜ 0.01) ; peripheral blood HBV-specific CTL levels were 0.65％±0.10％ and0.48％±0.07％,respectively (t=9.61,P＜0.01).Conclusions After treatment with ADV,along with the decrease of HBV DNA load,HBV-specific CTL surface PD-1 expression decreases,while HBV-specific CTL level increases.The changes in low virus load group are much more remarkable.

Objective To approach the application value of diffusion weighted imaging with background suppression (DWIBS) combined with routine MR sequence in differentiating benign and malignant lymph nodes,and assessing therapeutic effect of lympho-genic tumors.Methods 48 patients with cervical lymph node enlargement who were confirmed by pathology and follow up under-went DWIBS and routine MRI examination,malignant lymph nodes were 83,benign lymph nodes were 79.16 patients with malig-nant lymph nodes were rechecked after radiotherapy and chemotherapy,Apparent diffusion coefficient (ADC)values of the solid part for lymph nodes were compared.Results More lymph nodes can be detected in DWIBS than conventional sequence.ADC values of the solid part for malignant lymph node(0.898±0.111)×10-3 mm2/s were lower than that of benign lymph node(1.043±0.106)× 10-3 mm2/s,there was significant difference between them (P<0.05).ADC values of all malignant lymph nodes after treatment (1.205±0.121)×10-3 mm2/s were significantly higher than that of pretherapy (0.883±0.090)×10-3 mm2/s (P<0.05).Conclu-sion DWIBS could more sensitively detect lymph node than conventional MR sequence.ADC value could provide some reference values for differentiating benign and malignant lymph nodes and assessing therapeutic effect.

Objective To investigate the role of muhiplex ligation-dependent probe amplification(MLPA) in identifying fetal aneuploidy of chromosomes 13,18,21,X,and Y. Methods From June 2007 to December 2008,263 samples(prenatal diagnosis group),including amniotic or umbilical cord blood from pregnant women who required prenatal diagnosis,were processed in parallel by MLPA and conventional karyotype to detect fetal aneuploidy.Another 26 samples(fetal death group).ineluding retained abortion and fetal death,were also processed bv MLPA. Results Five cases of 21-trisomy,4 eases of 18-trisomy,1 case of 13-trisomy and 3 cases of 45,X were identified among the prenatal diagnosis group by MLPA,and the results were consistent with karyotype.Two cases of 45,X and 1 case of 18-trisomy were identified among the retained abortion and fetal death group. Conclusions MLPA is a rapid,efficient,simple,reliable and economical technique in detecting most common chromosomal aneuploidies and have important clinical value.