Objective To evaluate cytotoxic effects of cytokine-induced killer cells (CIK cells) transfected with the interleukin-2 (IL-2) gene on malignant melanoma cells.Methods Mouse spleen cells were extracted,lymphocyte cells were separated,and CIK cells were prepared from these lymphocyte cells.PEGF-N1 plasmids containing IL-2 gene (PEGF-NI-IL-2) were transfected into CIK cells.Fluorescence microscopy was used to observe transfection products,and reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to determine the IL-2 mRNA expression.Then,effector cells such as CIK cells and IL-2-transfected CIK cells were separately co-cultured with target cells (B16 melanoma cells) at effector-target ratios of 10∶ 1,20∶1 and 40∶1,then 4-hour lactate dehydrogenase release assay was performed to evaluate cytotoxic effects of the two kinds of CIK cells on B 16 cells.After effector cells were cocultured with target cells at the effector-target ratio of 40∶1 for 48 hours,enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-2,interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant of the two kinds of CIK cells.Finally,mouse models of melanoma were established,and a total of 28 melanoma-bearing mice were divided into 4 groups to be peritumorally injected with 0.2 ml sodium chloride physiological solution (control group),100 IU IL-2 solution (IL-2 group),CIK cell suspension at a cell density of 1 × 106 cells per milliliter (CIK group) and IL-2-transfected CIK cell suspension at a cell density of 1 × 106 cells per milliliter (IL-2-transfected CIK group) respectively.Tumor morphology,tumor inhibition rate and cell apoptosis rate were used to evaluate tumor growth in the above groups.If data were normally distributed,t-test was used for comparing means between two groups,and analysis of variance and least significant difference (LSD)-t test were used for comparing means among multiple groups.Results Fluorescence microscopy and RT-PCR both showed that IL-2 was successfully transfected into CIK cells.The cytotoxic effect of IL-2-transfected CIK cells on B16 cells was strongest at the effector-target ratio of 40:1.Levels of IL-2,IFN-γ and TNF-α were also significantly higher in the supernatant of IL-2-transfected CIK cells [(1107.26 ± 6.49) pg/ml,(50.01 ± 3.35) pg/ml,(39.86 ± 3.25) pg/ml] than those in that of CIK cells [(51.09 ± 3.85) pg/ml,(32.71 ± 2.43) pg/ml,(30.11 ± 3.08) pg/ml,t =442.60,14.93,6.89,all P ＜ 0.01].Animal experiments showed that the tumor volume obviously increased in the control group (P ＜ 0.05),but significantly decreased in the IL-2 group,CIK group and IL-2-transfected CIK group (all P ＜ 0.001) after intervention compared with those before intervention.Furthermore,the tumor volume in the IL-2-transfected CIK group was significantly less than that in the other three groups (all P ＜ 0.01),but no significant difference was observed between the IL-2 group and CIK group (P ＞ 0.05).In addition,the apoptosis rate was significantly higher in the IL-2 group,CIK group,and IL-2-transfeeted CIK group than that in the control group (all P ＜ 0.01).The apoptosis rate and tumor inhibition rate were significantly higher in the IL-2-transfected CIK group than those in the IL-2 group and CIK group (all P ＜ 0.01),but insignificantly different between the IL-2 group and CIK group (P ＞ 0.05).Conclusion IL-2-transfected CIK cells had stronger killing effects on malignant melanoma.

Objective Ti explire the effect if clinical nursing pathway in elderly fracture patients.Methods 60 senile femiral intertrichanteric fracture patients were divided inti the ib-servatiin griup and the cintril griup,with 30 patients in each griup.The cintril griup was given riutine nursing while the ibservatiin griup was given clinical nursing pathway.Recivery and life quality if the twi griups were cimpared after iperatiin.Results Health kniwledge scire,reha-bilitatiin enthusiasm,cimpliance in the ibservatiin griup was significantly higher than that in the cintril griup and the pistiperative cimplicatiins were significantly less than that if the cintril griup.The hispitalizatiin time was significantly shirter than that in the cintril griup and nursing satisfactiin was significantly higher than in the cintril griup.Thus,the difference was statistical significance (P <0.05).The difference was statistical significance since the life quality scire in health,physical functiin,bidy pain in the ibservatiin griup were significantly higher than thise in the cintril griup (P <0.05).Conclusion Clinical nursing pathway imprives the enthusiasm if elderly fractures patients,reduces the pistiperative cimplicatiins and imprives the quality if life fir patients.

Objective Ti explire the effect if clinical nursing pathway in elderly fracture patients.Methods 60 senile femiral intertrichanteric fracture patients were divided inti the ib-servatiin griup and the cintril griup,with 30 patients in each griup.The cintril griup was given riutine nursing while the ibservatiin griup was given clinical nursing pathway.Recivery and life quality if the twi griups were cimpared after iperatiin.Results Health kniwledge scire,reha-bilitatiin enthusiasm,cimpliance in the ibservatiin griup was significantly higher than that in the cintril griup and the pistiperative cimplicatiins were significantly less than that if the cintril griup.The hispitalizatiin time was significantly shirter than that in the cintril griup and nursing satisfactiin was significantly higher than in the cintril griup.Thus,the difference was statistical significance (P <0.05).The difference was statistical significance since the life quality scire in health,physical functiin,bidy pain in the ibservatiin griup were significantly higher than thise in the cintril griup (P <0.05).Conclusion Clinical nursing pathway imprives the enthusiasm if elderly fractures patients,reduces the pistiperative cimplicatiins and imprives the quality if life fir patients.

OBJECTIVETo evaluate the influence of small-dose dexmedetomidine infusion on recovery of patients undergoing vertebral operation.

METHODSSixty ASA I-II patients undergoing vertebral operation were randomly divided into two groups (n=30). In group I, dexmedetomidine infusion was pumped at the rate of 0.5 µg·kg(-1)·h(-1) from tracheal intubation to incision suture, and in group II, 0.9%saline was pumped instead. The mean arterial pressure, heart rate, Riker Sedation-Agitation Scale and Ramesay sedation score were recorded at the time points of autonomous respiration (T1), eye opening (T2), extubation (T3), 1 min after extubation (T4), 10 min after extubation (T5), and 30 min after extubation (T6).

RESULTSThe recovery time of autonomous respiration and eye opening time in group I were significantly longer than those in group II, and the extubation time was significantly shorter in group I (P<0.05). Riker Sedation-Agitation Scale scores in group II were significantly higher than those in group I at T2 and T4, and Ramesay sedation scores in group I were significantly higher than those in group II at T1, T2 and T5 (P<0.05). The mean arterial pressure and heart rate at each time point was significantly lower in group I than in group II (P<0.05), especially at T3 and T4 (P<0.01). In both groups, the mean arterial pressure and heart rate at T3 and T4 were significantly higher than those at rest (P<0.05).

CONCLUSIONSSmall-dose dexmedetomidine infusion can reduce dysphoria and lower the risks during recovery from general anesthesia following vertebral operation.

Adult ; Aged ; Anesthesia Recovery Period ; Dexmedetomidine ; administration & dosage ; pharmacology ; Humans ; Middle Aged ; Spine ; surgery

Objective To analyze problems existing in our scientific research methodological service process in our hospi-tal and realize the continuous improvement of service quality through project management .Methods We took the method of PDCA cycle management model and combined with the project management tools (such as fishbone diagram brainstorming method was used to perform causal analysis ,adopting the Gantt chart making project plan ,using SMART principle to set the target theme ,etc .) ,by using some strategies such as improving the standard of service system for departments ,establishing a professional and partitioned service flow ,adjusting the division of labor according to professional specialties ,communicating and learning to optimize solutions regularly ,equipping with professional technicians to improve operation and maintenance secu-rity of service platform ,etc .Results We completed the three target themes of service quality improvement about "shortening the period of service statistics analysis report","improving the accuracy of scientific research design"and"reducing the number of service data platform problem report".Conclusions The adoption of PDCA cycle management can effectively improve the quality of scientific research methodological service in the hospital .

Objective To evaluate the safety of cetuximab combined with intensity-modulated radiotherapy (IMRT) plus concurrent cisplatin chemotherapy in locoregionally advanced nasopharyngeal carcinoma (NPC) in a Chinese multicenter clinical study.MethodsFrom July 2008 to April 2009,100Patients with primary stage Ⅲ- Ⅳb non-keratinizing NPC were enrolled.The planned dose of IMRT to gross tumor volume and positive cervical lymph nodes was 66.0-75.9 Gy and 60-70 Gy in 30-33 fractions.Cisplatin (80 mg/m2,q3 week (w)) and cetuximab (400 mg/m2 one w before radiation,and then 250mg/m2 per w) were given concurrently.The adverse events (AEs) were graded according to common terminology criteria for adverse events v3.0.ResultsThe compliance of the entire group of patient was satisfactory.Actual median dose to gross tumor volume was 69.96 Gy,and the median dose to positive cervical lymph nodes was 68 Gy.Median dose of cisplatin was 133 mg,median first-dose of cetuximab was 690 mg,and median weekly dose was 410 mg.AEs were well tolerated and manageable,mainly consisting of acneiform skin eruptions,dermatitis and mucositis.Grade 4 mucositis was observed in 2％ of the patients and no other grade 4 AEs were observed.ConclusionsThe combined treatment modality of IMRT +concurrent chemotherapy + cetuximab in loco-regionally advanced NPC is well tolerated.

Objective: To explore the synergistic effect of silibinin combined with crizotinib on anaplastic lymphoma kinase positive (ALK+ ) non-small cell lung cancer (NSCLC) cells and its mechanism. Methods: H2228 and H3122 cells were treated with silibinin, crizotinib alone or in combination. Cell proliferation was measured by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation assay. Migration or invasion ability was tested by wound healing assay or transwell assay, respectively. Expressions of E-Cadherin and vimentin protein were examined by immunofluorescence staining. The protein expressions of ALK, p-ALK, E-Cadherin and Vimentin were detected by western blotting.The anti-cancer effect of silibinin combined with crizotinib in vivo was determined by subcutaneously injecting 2×10⁶ H2228 cells into immunodeficient nude mice. Results: The result of MTT assay showed that the cell viability of H2228 or H3122 treated with 100 μmol/L silibinin was (88.38±4.10)% or (72.27±3.62)%, respectively, marginally decreased compared with that of the control. The 50% inhibitory concentration (IC₅₀) of H2228 cells treated with crizotinib alone or combined with 100 μmol/L silibinin was (917.10±7.75) nmol/L or (238.73±7.67) nmol/L, respectively. The IC₅₀ of H3122 cells treated with crizotinib alone or combined with 100 μmol/L silibinin was (472.50±15.70) nmol/L or (206.10±12.01) nmol/L, respectively. The IC_50s of H2228 and H3122 cells were significantly decreased by combined treatment of crizotinib and silibinin compared to crizotinib treatment alone (P<0.01). When compared with the control group, colony forming ratios of H2228 cells were (83.34±2.72)% in 100 μmol/L silibinin treatment group, (69.42±3.06)% in 400 nmol/L crizotinib treatment group and (27.32±1.42)% in combined treatment group. When compared with the control group, colony forming ratios of H3122 cells were (84.45±5.67)% in 100 μmol/L silibinin treatment group, (45.02±5.83)% in 400 nmol/L crizotinib treatment group and (17.43±3.83)% in combined treatment group. Silibinin combined with crizotinib treatment significantly inhibited the colony formation ability of H2228 and H3122 cells (P<0.01). Migration and invasion results showed that combined treatment of crizotinib and silibinin markedly inhibited the migration and invasion ability of H2228 cells (P<0.01). Western blot results indicated that treated with silibinin alone or in combination of crozitinib for 48 hours, the protein level of E-cadherin in H2228 cells was upregulated, while the expressions of p-ALK and vimentin were downregulated, without obvious alteration of ALK protein expression. In the xenograft model, the mean tumor weight was (9.40±2.58)g in crizotinib treatment group and (4.58±1.07)g in the combined treatment group. The inhibitory effect of tumor growth in vivo of combined treatment was significantly superior to that of crizotinib treatment alone (P<0.05). Conclusion Silibinin enhances the inhibitory effect of crizotinib on ALK positive NSCLC cells, which may be associated with suppression of ALK activity and mesenchymal-epithelial transition.

Lung cancer is the leading cause of cancer death worldwide as well as in China. For many years, conventional oncologic treatments such as surgery, chemotherapy, and radiotherapy (RT) have dominated the field of non-small cell lung cancer (NSCLC). The recent introduction of immunotherapy in clinical practice, led to a paradigm shift in lung cancer as in many other solid tumors. Recent pre-clinical and clinical data have shown RT may also modify antitumor immune responses through induction of immunogenic cell death and reprogramming of the tumor microenvironment. This has led many to reexamine RT as a partner therapy to immuno-oncology treatments and investigate their potential synergy in an exponentially growing number of clinical trials. Clinical trials combining radiotherapy and immunotherapy are attracting major attention, experts were invited to discuss frontier and controversial academic topics: (1) Recent developments of clinical synergy between radiation and immune checkpoint inhibitors (ICIs) in the treatment of NSCLC; (2) Will immunotherapy and radiotherapy increase the toxicity risk for cancer patients; (3) How to cope the mixed responses/disassociated responses phenomenon in checkpoint inhibition therapy to NSCLC with local ablative therapy; (4) Combining radiotherapy and immunotherapy in the treatment of NSCLC brain metastases.