Objective To evaluate the effect and mechanism of IL‐6 induced Gefitinib resistance in non small cell lung cancer (NSCLC) .Methods The sensitivity of cells to Gefitinib ,the invasion ability of cells and the expression of phosphorylated p‐mTOR was assessed by MTT assay ,Transwell assay and Western blot ,respectively .PC‐9psb388 stable over expressing human recombi‐nant IL‐6(hrIL‐6) cell line was established by transfecting PC‐9 cells with a lentivirus psb388 expressing IL‐6 and stable transfecta‐nts over‐expressing IL‐6 in human lung cancer cell line PC‐9 .The sensitivity of cells to Gefitinib ,the invasion ability ,expression of p‐mTOR were then detected .PC‐9/PC‐9psb388 xenografts were established and the expression of p‐mTOR and IL‐6 in tumor sec‐tions were then detected .Results The sensitivity of PC‐9 cells to Gefitinib was reduced by IL‐6 ,the invasion ability of PC‐9 cells and the expression of p‐mTOR was significantly increased with IL‐6 treatment .The sensitivity of PC‐9 cells to Gefitinib was promi‐nent higher in PC‐9psb388 cells ,while the invasion ability of PC‐9psb388 cells and the expression of p‐mTOR was higher than PC‐9 cells .The sensitivity to Gefitinib was improved and expression of p‐mTOR reduced in rapamycin‐treated PC‐9psb388 cells and IL‐6 stimulated PC‐9 cells .Tumor volume of PC‐9psb388 xenografts was significantly higher than that of PC‐9 cells .The expression of p‐mTOR and IL‐6 in tumor sections of PC‐9psb388 group were higher than that of PC‐9 group .Conclusion IL‐6 could elevate the expression of p‐mTOR to induce Gefitinib resistance in non small cell lung cancer (NSCLC) .

Aged, 80 and over ; Chordoma ; complications ; Humans ; Male ; Pharyngeal Neoplasms ; complications ; Respiration Disorders ; etiology

OBJECTIVE: To develop a method of HPLC/MS/MS to determine phenacetin and paracetamol in rat liver microsomal incubation system. METHODS: Samples were separated on XTerra MS C18 column, different ratios of methanol- 0.1% formic acid were used as the gradient eluent, and the flow rate was 0.2mL?min-1. The electrospray ion-quadrupole mass spectrometry and multiple reaction monitor were adopted to detect the concentration of phenacetin and paracetamol. RESULTS: The calibration curves were linear in the ranges of 45～9 000ng?mL-1(r=0.999 8)and 15.2～1 520ng?mL-1(r=0.999 6) for phenacetin and paracetamol respectively; The lowest limits of assay were 9ng?mL-1 and 10ng?mL-1.The average recoveries at three concentrations of phenacetin were (96.2?2.3)%～(98.3?2.4)% and those of paracetamol were (99.6?2.1)%～(100.2?2.6)%; RSD of the intra-day and inter-day were less than 5%. CONCLUSION: The method is rapid, sensitive and suitable for determination of phenacetin and paracetamol in rat liver microsomal incubation system.

Background and purpose:It has beenreported that miR-1284 is associated with gastric cancer lymph node metastasis in the research of microRNA microarray in human gastric cancer tissues. But the specific role of miR-1284 in gastric cancer has not been reported. The aim of this study was to investigate the effect of miR-1284 over-expression on the gene expression profiling and invasion/metastasis of human gastric cancer SGC-7901 cells. Methods:Gastric cancer SGC-7901 cells of LV-miR-1284 group were transfected with lentiviral vectors of miR-1284, cells of LV-NC-GFP group were transfected with lentiviral vectors without miR-1284, and cells of control group were not transfected with lentiviral vectors. The expression of miR-1284 was detected by the real-time fluorescent quantitative PCR. Differential expression genes were detected by the microRNA chip. Target genes of miR-1284 were predicted by the bioinformatics. Invasive ability was detected by the Transwell invasion assay. Metastasis ability was detected by subcutane-ously transplanted tumor model of nude mice.Results:Compared with LV-NC-GFP and control groups, the expressions of miR-1284 and 20 genes were up-regulated, and the expression of 17 genes was down-regulated in LV-miR-1284 group. One hundred and thirty-eight target genes of miR-1284 were predicted by the bioinformatics website. Compared with invasive cell number of LV-NC-GFP group (168.67±4.55) and control group (170.33±3.08), the ability of invasion ofcells was weakened in LV-miR-1284 group (70.00±2.37). Compared with the liver metastasis rate of LV-NC-GFP group (85.71%) and control group (85.71%), the ability of metastasis of cells was weakened in LV-miR-1284 group (14.29%). Conclusion:The ability of invasion and metastasis of SGC-7901 cells is suppressed by over-expression of miR-1284. The mechanism may be related to regulating the expression ofSUMO1 andJUNgenes.

AIM:To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism.METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer ( GC) tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS:Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples ( P<0.05 ) .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05).Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased (P<0.05), the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05).CONCLUSION:The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.

Objective To examine the effectiveness of HGF in blocking TGF-β1 induced α-SMA and extracellular matrix production in fibroblasts of the flexor tendon sheath. Methods Seven adult male New Zealand white rabbits (3.75-4.00 kg) were used for this study. Both of their front feet were sterilised and the middle digit flexor digitorum profundus tendon equivalents were identified and isolated. These specimens were used to establish primary cell cultures. Sheath fibroblasts were obtained from rabbit flexor tendons. After the cells reached confluence, cells were detached with trypsin/ethylenediamine tetra-acetic acid. All experiments were performed using the cells at the third passage. At 70% confluence the medium was supplemented with 5 ng/ml of TGF-β1 along with increasing doses of HGF (10-40 ng/ml). After 72 hours incubation, the productions of α-SMA were assayed by Western-Blot. The productions of collagen Ⅰ and fibronectin in supernatants culture were examined using ELISA. Results Evaluation of protein expression revealed that TGF-β1 markedly induced α-SMA expression in cultured rabbit flexor tendon sheath fibroblasts. TGF-β1 treated fibroblasts expressed 1.8-fold more protein compared to non-treated fibroblasts (P ＜ 0.05). However, simultaneous incubation of HGF significantly abrogated TGF-β1 induced α-SMA expression in a dose-dependent manner (P＜ 0.05). Treatment with TGF-β1 significantly stimulated collagen Ⅰ and fibronectin production in flexor tendon sheath fibroblasts (P ＜ 0.01). Remarkably, the addition of HGF reduced productions of all components induced by TGF-β1 in a dose-dependent manner (P ＜ 0.05). Conclusion HGF antagonizes TGF-β1 induced α-SMA, collagen Ⅰ, and fibronectin production in flexor tendon sheath fibroblasts. The findings provide a cellular and molecular basis for HGF's acting as a therapeutic agent for adhesions in flexor tendons.

Objective:To investigate the therapeutic effects of antibiotic articular cement spacer on shoulder joint infection in the elderly patients during stage-one operation.Methods:The data of 3 patients were analyzed retrospectively who had been treated at Department of Orthopaedics, The Sixth People's Hospital Affiliated to Shanghai Jiao Tong University for shoulder infection from May 2018 to December 2019. They were one man and 2 women with an average age of 65.3 years (from 64 to 67 years). One case of infection followed shoulder puncture, another proximal humeral fracture and another shoulder prosthesis replacement. All the 3 patients underwent radical debridement and implantation of antibiotic shoulder cement spacer in stage-one operation but no stage-two operation. The therapeutic effects were evaluated by American Shoulder and Elbow Surgeons (ASES) scoring, Quick Disabilities of the Arm, Shoulder, and Hand (Quick-DASH) scoring, Visual Analog Scale (VAS), and range of shoulder motion.Results:The 3 patients were followed up for 18 to 28 months (mean, 22.7 months). There was no recurrence of infection and the spacers were in good position. The microorganisms detected were Bifidobacterium brevis in one case and methicillin-resistant Staphylococcus aureus in 2 cases. At the final follow-up, the ASES scores averaged 54.4 (from 46.3 to 60.0), Quick-DASH scores 45.1 (from 40.8 to 50.0), VAS scores 2.3 (from 2 to 3), ranges of elevation 65.7 °, ranges of abduction 43.8° and ranges of external rotation 21.7°.Conclusion:For the elderly patients with shoulder infection, implantation of antibiotic shoulder cement spacer after radical debridement can well control infection, relieve pain and improve shoulder functions, sparing them secondary operation.

Objective To discuss the influence of grading nursing care by physician-nurse cooperation decision-making on the quality of patient care and disease outcomes,and evaluate the application feasibility of nursing core system in this nursing mode.Methods 379 patients were admitted to departments of orthopedics and endocrinology from two hospital areas of our hospital from January 2011 to January 2012.Patients were divided into two groups according to nursing process.Patients who were given conventional grading nursing were set as the routine group,and patients receiving physician-nurse collaboration grading nursing were set as the collaboration group.The nursing quality and nursing efficiency,and the disease outcome of patients were compared between the two groups.Results The quality of care,the qualified rate of care plan implementation,the coverage rate of health education of patients and the patient satisfaction rate were higher in the collaboration group than those in the routine group.The performing rate of doctor's orders,the awareness rate of patient disease,the average length of hospital stay,compliance of medical orders and re-admission rate of patients with the same or a related disease within three months were significantly different between two groups.The collaboration group improved the matching degree of nursing grade with disease condition,understanding of professional knowledge,recording of nursing records,deficient nursing process,and communication ability of life knowledge,compared with those of the routine group.Conchusions Nursing grading mode by physician-nurse collaboration decision-making can obviously improve the defmition of grading nursing and the general treatment effect of patients,it significantly improved the quality of grading nursing.

BACKGROUND:Cytotoxic T lymphocyte-associated antigen 4 is a newly discovered costimulatory molecule. It has been studied more in tumor and autoimmune diseases, less in the field of kidney transplantation.
OBJECTIVE:To explore the role of cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation.
METHODS:Fifty patients undergoing renal transplantation were divided into acute rejection group (20 cases) and stable graft function group (30 cases). Another 30 healthy persons served as control group. Blood samples were extracted from the peripheral blood. Cytotoxic T lymphocyte-associated antigen 4 was detected by enzyme linked immunosorbent assay and flow cytometry.
RESULTS AND CONCLUSION:The expression of cytotoxic T lymphocyte-associated antigen 4 in the serum showed significant differences in the acute rejection group, stable graft function group and healthy control group (F=70.008 1, P=0.000 0), but showed no difference in peripheral blood lymphocytes of three groups (F=1.865 6, P=0.161 7). Compared with the healthy control group, the expression levels of cytotoxic T lymphocyte-associated antigen 4 in peripheral blood lymphocytes of acute rejection group and stable graft function group were significantly decreased (P=0.000 0). In addition, the acute rejection group had a lower cytotoxic T lymphocyte-associated antigen 4 expression than the stable graft function group (P=0.000 0). In renal transplant rejection, the expression of cytotoxic T lymphocyte-associated antigen 4 in serum was reduced, showing some correlation with acute rejection after renal transplnatation. Cytotoxic T lymphocyte-associated antigen 4 might be involved in the rejection.

Objective:To investigate the influence of endoplasmic reticulum stress (ERS) in the degeneration of cochlear hair cells in the type 2diabetic mice, and to clarify its mechanism.Methods:Twenty clean Kun Ming male mice aged one month were selected and randomly divided into control group and model group (n=10) .The mice in model group were injected with STZ (40 mg·kg-1) to establish the type 2diabetic models.The fasting blood glucose levels of the mice were measured through collecting the vena caudalis blood of the mice.Auditory brain stem response (ABR) was used to detect the ABR threshold of the mice.Otoacoustic emission (OAE) test was used to detect the OAE threshold of mice.The defect rate of mouse cochlear outer hair cells was calculated by the mouse cochlear spreading technique.The expression levels of GRP78, caspase-12, p-ERK and Nrf2proteins were detected by Western blotting method.Results:Compared with control group, the fasting blood glucose levels of the mice in model group at the 7th and the 14th days had no significant differences (P＞0.05) , but the levels were increased significantly at the 21th, 28th and 35th days and the level reached the highest value at the 35th day.The ABR thresholds of the mice in model group at 8, 12, and 24kHZ were increased significantly compared with control group (P＜0.05) .Under the stimulation of low frequency, there was no significant change in the OAE threshold of the mice in model grouop compared with control group.The OAE thresholds of the mice in model group were increased significantly under the medium frequency and high frequency stimulation compared with control group (P＜0.05) .The defects of the cochlear hair cells were mainly concentrated on the bottom of gyrus of the mice, and the defects in middle temporal gyrus and parietal gyrus were less.Compared with control group, the defect rate in the bottom of gyrus of the mice in model group was increased significantly (P＜0.05) ;the defect rates in the middle temporal gyrus and parietal gyrus were increased, but there was no significant difference (P＞0.05) .The expression levels of p-ERK and Nrf2in the cochlear hair cells of the mice in model group were lower than those in control group (P＜0.05) , and the expression levels of GRP78and caspase-12were higher than those in control group (P＜0.05) .Conclusion:ERS can result in the increase of defect rate of cochlear outer hair cells and ABR brainstem hearing threshold of the diabetic mice and decrease the expression levels of p-ERK and Nrf2proteins, suggesting that ERS can promote the degenerative lesions of cochlear hair cells in the type 2diabetic mice.