Objective MiRNA-10b is an important member of the MiRNA family , which has been proven that miRNA-10b can promote the growth and invasion of a variety of tumor cells .The aim of this study was to to investigate the effect and mechanism of miR-NA-10b on proliferation and invasion of low metastasis of lung cancer cell line (95-C). Methods The recombinant of miRNA-10b was transfected into 95-C by lipofectin method .The experiment set up 3 groups:blank control group , negative control group and miRNA-10b expression plasmid transfected group .MiRNA-10b expression level and KLF4mRNA expression level were detected by real-time fluores-cence quantitative PCR ( RTFQ-PCR) .The cell proliferation was detected by cell proliferation assay .The invasive ability of cell was de-tected by Transwell experiment .The expression of KLF4 protein was assessed by Western blot . Results At 48 hours after transfection, compared with blank control group (1.01 ±0.08) and negative control group (0.86 ±0.07), the miRNA-10b expression level in miRNA-10b expression plasmid transfected group (1.61 ±0.12) increased significantly (P<0.05) and there was no statistical difference between blank control group and negative control group (P>0.05).From the growth curve, the cell proliferation rate was obviously increased in miRNA-10b expression plasmid transfected group ([188.0 ±15.1]/HP) compared with the other two groups ([151.0 ±11.3]/HP), ([136.0 ±10.8]/HP) (P <0.05) and there was no statistical difference between these two groups ( P >0.05 ).Transwell result showed more cells transferred to the other side of Transwell compared with the other two groups ( P <0.05 ) and there was no statistical difference between these two groups (P >0.05).The expression of KLF4 protein decreased in miRNA-10b expression plasmid transfected group compared with the other two groups ( P<0.05).KLF4mRNA expression decreased, but the difference had no statistical significance (P>0.05). Conclusion MiRNA-10b might promote the pro-liferation and invasion of 95-C through down-regulation of KLF4 protein expression .

Objective To analyze the clinical features of juvenile dermatomyositis (JDM) combined with soft-tissue calcification. Methods Forty-seven patients with JDM combined with soft-tissue calcification (soft-tissue calcification group) were retrospectively analyzed, and they were contrasted with 89 patients with non-calcification (non-calcification group). Results The rates of Gotton signe, muscle contracture and joint dysfunction in soft-tissue calcification group were significantly higher than those in non-calcification group:87.23% (41/47) vs. 43.82% (39/89) and 68.09% (32/47) vs. 21.35% (19/89), and there were statistical differences (P<0.05). The dosage of glucocorticoid (conversion of prednisone measuring more than 1.5 mg/kg), rate of using immunodepressant, level of creatine kinase in soft-tissue calcification group were significantly lower than those in non-calcification group:17.02%(8/47) vs. 68.54%(61/89), 25.53%(12/47) vs. 88.76%(79/89), (566.45±240.41) U/L vs. (1 680.12±656.50) U/L, and there were statistical differences (P<0.05). Conclusions The patients with JDM combined with Gotton signe are more prone to soft-tissue calcification. The rate of muscle contracture and joint dysfunction in soft-tissue calcification patients is significantly higher than that in non-calcification patients. For the patients whose creatine kinase are not obviously elevated, they are more prone to soft-tissue calcification. Early active application of glucocorticoid and immunodepressant therapy can reduce or prevent the occurrence or development of late calcium deposition.

ObjectiveTo assess the feasibility and efficacy of a docetaxel plus cisplatin regimen for patients of locally advanced nasopharyngeal carcinoma(NPC)treated concurrently with definitive IMRT in a short-term observation.MethodsRadiation consisted of 7000 cGy given to the planning target volume (PTV) of primary tumor, 6600 cGy given to the PTV of metastatic lymph nodes and 6000 cGy to the PTV of subclinical disease in 220-228 cGy/fraction were delivered over 31-32 treatment days. Thirty-two patients with newly diagnosed NPC received definitive intensity-modulated radiation therapy(IMRT)concurrent with docetaxel 75 mg/m2 on day 1 and DDP 75 mg/m2 on day 1(or DDP 25 mg/m2 on day 1-day 3), repeating every 21 to 28 days for 2 cycles.ResultsAll patients received the full dose of radiotherapy and completed 2 cycles of chemotherapy with a median follow-up of 13 months (2-28 months).No treatment-related death was observed. Major toxicities included hematologic toxicity and mucositis. The incidence rates of grade 3-4 leucopenia,grade 3-4 neutropenia and grade 3 acute mucositis were 46.9 ％ (15/32),59.4 ％ (19/32) and 40.6 ％ (13/32) respectively.The complete remission (CR) rate was 96.9 ％ (31/32).During treatment,90.6 ％ (29/32)patients acquired granulocyte colony stimulating factor (G-CSF)for leucopenia. The 1-year overall survival, local recurrence-free survival, regional recurrence-free survival and distant metastasis-free survival were 100 ％ (31/32),96.9 ％ (31/32),96.9 ％ (31/32),96.9 ％ (31/32),respectively,for the whole cohort.Conclusions2 cycles of the docetaxel plus cisplatin regimen with concurrent IMRT are demonstrated being feasible and effective in treating locally advanced NPC with promising results.The major toxicities are leucopenia and neutropenia, but they are tolerable with the use of G-CSF. Further investigation of long-term efficacy of the regimen is required.

Objective By culturing the osteoclasts together with the osteoblasts directly to investigate the effect of osteoblasts on the formation of mature osteoclasts.Methods The bone marrow mononuclear cells of rats were treated with 30?g/L M-CSF and 50?g/L RANKL and cultured for 6 days.Subsequently,the primary osteoblasts which were of the same quantity as the osteoclasts were co-cultured directly.In the co-culture system,we added the liquid containing 1,25-(OH)2D3 1?10-8mol/L and PGE2 1?10-6mol/L.The morphological observation,TRAP staining and pit staining were adopted to identify osteoclasts.Results When the osteoclasts were co-cultured with primary osteoblasts,the growth of osteoblasts had more preponderances.After staining,we could see more osteoblasts than osteoclasts.Conclusion The relationship between osteoblasts and osteoclasts is related to the relative quantities of the two cells.When osteoblasts outnumber osteoclasts,osteoblasts would inhibit the formation and differentiation of osteoclasts.

Objective To compare the displayed inner diameter of coronary stent by high definition(HD)and gem?stone spectral imaging(GSI)using dynamic cardiac and coronary artery phantom. Methods Five different types of coro?nary stents(internal diameter, 3.10 mm±0.55 mm;strut thickness, 0.12 mm±0.04 mm)were placed into a pulsating cardiac phantom(ALPHA 1-VT PC, Fuyo Corporation, Japan). The stent phantom was scanned by 3 systems, gemstone spectral im?aging(GSI), spiral scan(S)and HD. All the spectral imaging data were analyzed using GSI viewer to reconstruct the VMS (monochromic spectral) images(60-140 keV). Image noise(N), signal-to-noise ratio(SNR), contrast-to-noise ratio(CNR) and inner diameter were compared between images acquired through these 3 systems. Results SNRs in images of S and HD were higher than that of GSI(P<0.05),while there were no significant differences in CNRs among images of GSI, S and HD(P>0.05). The visible diameter(%)measurements of HD(0.85 ± 0.06)was significant higher than that of the other 2 scan systems and most close to the width of those stent’s actual size.(GSI:0.40±0.16, 0.48±0.13, 0.50±0.14, 0.51±0.13, 0.45±0.05,0.52±0.13, 0.53±0.13, 0.53±0.13, 0.53±0.13, S:0.53±0.14, P<0.05). Conclusion There was no significant dif?ferences in image quality among the images acquired by these 3 systems when the heart rate was set to 60 beats per min. Comparing to GSI and S, HD can produce best represent images to the known inner diameter of coronary stent.

OBJECTIVE To prepare pneumolysin(Pn)by genetic engineering and thereby establish the basis for the study of vaccines against otitis media. METHODS A pair of primers including two restriction sites was designed based on the pneumolysin gene sequence reported by Walker in 1987. The pneumolysin gene was PCR-amplified from pneumococcal DNA. The resulting fragment, digested by restriction enzymes, was ligated into the vector PET-28a and then transformed into host cell E.coli JM109(DE3). RESULTS The sequence of the inserted pneumolysin gene was confirmed by DNA sequencing. CONCLUSION The pneumolysin gene was successfully cloned into the host cell.

A HPLC method for simultaneous determination of uridine, inosine, guanosine, adenosine and cordycepine
in Cordyceps sinensis was developed. The sample solution was prepared with 0.5% phosphoric acid solution by ultrasonic extraction. The separation was performed on a ZORBAX SB-AQ (150 mmí4.6 mm, 5 μm)column with gradient elution by 0.1% formic acid solution and acetonitrile, with column temperature 30℃, at a flow rate of 0.8 mL/min, and detected at wavelength of 260 nm. The result of method validation showed that the developed method had high accuracy and good repeatability. This method has been successfully applied for analysis of 5 kinds of nucleosides in different parts of C. sinensis. The results indicated that content of nucleosides in stroma is higher than that in insect body and whole C. sinensis.

A HPLC-QTOF MS method was established for analysis of components in Ophiocordyceps sinensis . The HPLC analysis was performed on an Agilent Zorbax SB Aq (150 mmí4.6 mm, 5 μm) with gr adient elution (5 mmol·L-1 ammonium acetate aqueous solution-acetonitrile), flow rate was 0.8 mL·min-1 and detection wave-length was 260 nm. The developed method was successfully applied in analysis of three different samples in-cluding O. sinensis, Hirsutella sinensis ( anamorph of O.sinensis) and Cordyceps militaris. Nine compounds were i-dentified in both O.sinensis and H.sinensis, which eight compounds were identified in C.militaris.

Objective To observe diffusion changes of epiphysis of femoral head with ischemia of difference phases by line-scan diffusion weighted imaging(LSDWI),and determine whether LSDWI can provide temporal information and severity about ischemia of epiphysis.Methods lschemia was surgically induced in one hip of each piglet(n=25)and the other hip served as a normal control.Piglets were imaged before surgery and at 3 hours,72 hours and 1,3 and 6 weeks after surgery by using LSDWI.Apparent difrusion coefficients(A DC)in epiphysis of the femoral heads were calculated.Significant difierences in ADC values between ischemia group and control group were found by using paired t-test.After scan at individual time points,5 piglets were sacrificed for histological study each time.Results The ADC value in the ischemic femoral heads f(1.22±0.37)×10-3 mm2/s]decreased significantiy at 3 hours after surgery (t=3.914,P＜0.01),compared to that in control[(1.73±0.33)×10-3mm2/s},and increased at 72 hours[(2.15±0.32)×10-3mm2/s versus(1.70±0.22)×10-3 mm2/s](t=3.348,P＜0.01).Then ADC valne kept increasing until 6 weeks after surgery[(1.61±0.27)×10-3mm2/s in ischemia side vs (1.11±0.45)×10-3mm2/s in the control](t=4.136,P＜0.01).rrhe percentage change of the ADC value significandy increased at 3 hours,72 hours,1 and 3 week(s)after the surgery(P＜0.01),compared to that at the prior neighboring time point.No significant increase in the percentage change of ADC value was found between the 3rd week and the 6th week after the surgery(t=2.29.P＞0.05).Histological examinations revealed abnormal thickening within epiphyseal cartilage,and cartilaginous islands within ossified tissues.Growth disturbante wag found in form of focal growth plate disruption.Conclusions Dynamic changes of ADC values were found with the prolonged ischemia of the femoral head by LSDWI.It could serve as a useful marker for evaluating duration and extent of ischemic epiphyseal disruption.

Objective:To explore the expressions of endogenous hydrogen sulfide (H2 S)and its synthases cystathionine beta synthase (CBS)and cystathionine gamma lyase (CSE)in the cell lines of normal bladder and bladder cancer,and to clarify their mechanism in the development of bladder cancer.Methods:The bladder cancer cell lines (5637,T24,UM-UC-3,EJ)and human bladder epithelial cell line SV-HUC-1 were selected.The expressions of CBS and CSE in bladder cancer and normal cell lines were analyzed by Western blotting assay and the productivities of H2 S in cell lines were detected by sensitive sulphur electrode assay.The EJ cells were selected based on the previous experimental results and divided into groups as follows:① 10 μmol· L-1 NaHS group, 50 μmol·L-1 NaHS group,100 μmol·L-1 NaHS group and control group.After drug treatment,the cell survival rate was measured by MTT assay at 24 and 48 h.② 5 μg·L-1 cisplatin group,cisplatin (5 μg·L-1 )+ NaHS (100 μmol·L-1 )group and control group.After medicine treatment,the cell survival rate was measured by MTT assay and the cell apoptotic rate was detected by flow cytometry at 48 h. Results:Compared with the normal bladder cells (SV-HUC-1),the expression levels of CBS and CSE and the productivity of H2 S in the bladder cancer cell lines (5637,T24,UM-UC-3 and EJ)were increased obviously (P <0.05 or P <0.01).Compared with control group,exogenous H2 S promoted the cell proliferation of EJ cells.The cell survival rates were increased with the increase of drug dose (P <0.05),which showed a dose-dependent effect.The cell survival rates were increased with the prolongation of time (P <0.05),which showed a time-dependent effect.After medicine treatment,compared with cisplatin group,the cell viability in cisplatin+NaHS group was increased (P <0.05)and the apoptotic rate was decreased (P <0.05).Conclusion:Endogenous H2 S and its synthases CBS and CSE have an increased expression level in bladder cancer cell lines compared with the normal bladder cells.H2 S can enhance the proliferation of bladder cancer cells and decrease the apoptosis induced by cisplatin.