Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.

Objective To introduce the method and experience of the setting of psychological care nurses,to discuss its effect on high-quality nursing and on amelioration of the patients' negative moods.Methods Psychological care nurses were set up in all the clinical departments,after systemic training of psychological knowledge and became qualified psychological consultant via national certification examination.We took psychological care nurses as the leading role,assisted with other nursing modes.By the means of mental status scale in non-psychiatric settings (MSSNS) and the satisfaction questionnaire of psychological care to evaluate the patients' mental status and satisfaction degrees before and after the intervention in 297 cases.Results Patients'scores of anxiety scale,depression,anger,loneliness factor and the total score of the MSSNS after the psychological nursing intervention were significant differences than those before the psychological nursing intervention,[(39.66±4.70)vs (45.59±6.45),(23.31±2.28)vs (26.59±3.35),(13.91 ±3.01) vs (15.12±2.58),(17.58±3.74)vs (20.29 ±4.67),(92.09±6.85)vs (106.66±8.58),t values was 14.376,15.213,23.992,13.974,13.641 respectively,P＜0.05].The satisfaction rate of patients for psychological nursing intervention after the intervention was 96.97％(288/297),which was significant higher than that before the intervention,69.70％(207/297),x2=79.53,P＜0.05.Conclusions Psychological care demands professional qualities.Only through systemic training,can grasp the nurses master professional psychological knowledge and skill,so as to apply scientific psychological care to the patients and help them to solve their psychological perplexities and maintain physical and psychological well-beings.

ObjectiveTo evaluate the effectivenes of continous passive motion(CPM) on limb function in patients after total knee arthroplasty.MethodsThe Cochrane Database of Systematic Reviews, MEDLINE, EMBASE, PEDro, CNKI and VIP were retrieved from the date of establishment of the databases to July, 2010 to collect randomized controlled trials(RCTs). The quality of RCTs was critically appraised and data were extracted by 4 reviewers independently. Meta-analysis was conducted for the eligible RCTs.Results26 RCTs were included. The result of Meta-analysis showed that CPM improved the range of motion (ROM) in active knee flexion and passive knee flexion, eliminate swelling in short term(＜6 weeks) and alleviate pain in short and medium term (less than 6 months). CPM showed no significant difference in improving the ROM in active and passive knee extention and quadriceps strength, and shortening hospital stay.ConclusionThe patients carried out CPM training after total knee arthroplasty can effectively improve limb flexion function, eliminate joint swelling, and relieve pain in the short term. But more high-quality and large-scale RCTs are needed.

OBJECTIVETo observe the effect of Tetramethyl pyrazine (TMP) on the cytokines and inflammatory mediators in the serum and the synovial fluid of collagen-induced arthritis (CIA)rats, and further to investigate its possible mechanisms for treating rheumatoid arthritis (RA).

METHODSType II CIA rat model was established. Rats in the TMP group were administered with TMP at 50 mg/kg and 100 mg/kg, once daily. Dexamethasone at 2.0 mg/kg was intramuscularly injected to those in the Dexamethasone treated group, once daily. Normal saline at 2 mL/kg was given to those in the normal control group and the model group, once daily. All medication was started from the 7th day, lasting to the 35th day. CIA rats' foot swelling degree was observed. Contents of serum IL-1, IL-6, IL-2, NO and PGE2in the synovial fluid were detected by radioimmunoassay and nitrate reduction method.

RESULTSCompared with the normal group, the foot swelling obviously increased, contents of NO and PGE2 in the synovial fluid were obviously elevated in the model group (P < 0.01). Compared with the model group, the foot swelling could be obviously inhibited by 100 mg/kg TMP and Dexamethasone; serum levels of IL-1 and IL-6 obviously decreased, serum IL-2 level obviously increased, contents of NO and PGE, decreased (P < 0.01). TMP 50 mg/kg could obviously inhibit the foot swelling of CIA rats (P < 0.01). There was no statistical difference in other indices (P > 0.05).

CONCLUSIONSTMP at 100 mg/kg showed obvious inhibition on CIA rats. Its inhibitory effect might be correlated to inhibiting activities of endogenous cytokines and the generation of inflammatory mediators in inflammation local regions, improving contents of anti-inflammation cytokines, and inducing the balance of the inflammatory cytokine network.

Animals ; Arthritis, Experimental ; blood ; metabolism ; Dinoprostone ; metabolism ; Female ; Interleukin-1beta ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Nitric Oxide ; metabolism ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Synovial Fluid ; metabolism

Objective To test serum differentially expressed proteins between early-stage (stage IB-ⅢA) and late-stage (stage Ⅳ) lung cancer patients by proteinchip technology and investigate its clinical value. Methods SELDI-TOF-MS and WCX-2 protein chip were used to detect the serum protein of 30 cases of early stage lung cancer patients and 30 cases of late stage lung cancer patients. The data were analyzed by using Biomarker Wizard software. Results There are ten different proteins in the serum between the two groups of lung cancer patients. Four protein markers 7978, 8139, 15 951 and 16 133 are over expressed and seven protein markers 2867, 6885, 8701, 8840, 13 781 and 13 955 are low expressed in the late group. Conclusion SELDI-TOF-MS proteinchip technology is a convenient, sensitive and high-throughput analysis method which can screen several relatively specific protein markers for late stage lung cancer from the serum samples. This selected protein markers can predict metastasis of lung cancer patients.

OBJECTIVETo observe the effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.

METHODThirty-two NOD mice were randomly divided into the model group, the traditional Chinese medicine group (TCM group, orally given 0.4 mL nourishing Yin, strengthening Qi and activating blood decoction as per 100 g x kg(-1) everyday), the hydroxychloroquine group (given 0.4 mL hydroxychloroquine as per 60 mg x kg(-1) everyday), the traditional Chinese medicine and western medicine group (TCM WM group, given nourishing Yin, Strengthening Qi and activating blood decoction 50 g x kg(-1) and hydroxychloroquine 60 mg x kg(-1), 0.4 mL everyday), with eight mice in each group. Eight Balb/C mice were selected as the normal control group (normal group). All of mice were killed after eight weeks, and their submaxillary glands were dissected. The expression levels of Fas/FasL were examined by immunohistochemical method, and the FasL mRNA was detected by RT-PCR.

RESULTThe expression levels of Fas/FasL in salivary glands of the model group were higher than that of other groups (P < 0.05). The expression level of FasL of the normal group was much lower than that in the hydroxychloroquine group (P < 0.05). The relative expression level of Fas mRNA in salivary glands of the model group was higher than that in other groups, but the control group was notably lower than other groups (P < 0.05). The expression level of FasL mRNA in salivary glands of the model group was higher than that in TCM and TCM WM groups (P < 0.05). But the expression level in TCM WM group was notably lower than the hydroxychloroquine group (P < 0.05).

CONCLUSIONThe nourishing Yin, strengthening Qi and activating blood decoction could down-regulate the expression level of Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression, and had a better efficacy after being combined with hydroxychloroquine. The nourishing Yin, strengthening Qi and activating blood decoction might treat the Sjogren's Syndrome by reducing apoptosis which is regulated by Fas/FasL

Animals ; Fas Ligand Protein ; genetics ; Female ; Gene Expression Regulation ; Medicine, Chinese Traditional ; methods ; Mice ; Mice, Inbred NOD ; Qi ; RNA, Messenger ; genetics ; metabolism ; Salivary Glands ; metabolism ; Sjogren's Syndrome ; blood ; genetics ; therapy ; Yin-Yang ; fas Receptor ; genetics

Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.

Objective To prospectively compare the clinical efficacy between high versus low viscosity bone cement in percutaneous vertebroplasty (PVP) for severe osteoporotic vertebral compression fractures (OVCF).Methods A prospective study was conducted in 61 old patients with single severe OVCF who had sought medical attention in our hospital from August 2014 to October 2015.They were randomly assigned to group H (n =30) to receive PVP using high viscosity bone cement and group L (n =31) to receive PVP using low viscosity bone cement.The 2 groups were compared preoperatively and postoperatively in terms of visual analogue scale (VAS),Oswestry disability index(ODI),short Form-36 General Health Survey(SF-36),Kyphosis cobb's angle,height of the injured vertebra,and volume and leakage of bone cement.Results Significant improvements in VAS,ODI and SF-36 score were noted after operation in both groups,but there were no significant differences between the 2 groups (P ＞ 0.05).The 2 groups made significant improvements after operation in cobb's angle and height recovery rate of the injured vertebra;improvements in group H (14.7° ± 3.4° and 28％ ± 8％) were significantly greater than in group L (16.5° ± 2.5° and 22％ ± 7％) (P ＜ 0.05).The bone cement volumes in groups H and L were 4.94 ± 0.72 mL and 4.89 ±0.75 mL respectively,showing no statistically significant difference between the 2 groups (P ＞ 0.05).The leakage rate in group H was significantly lower than that in group L (13.33％ versus 35.48％) (P ＜ 0.05).All the patients were followed up for a mean time of 12 months (range,from 3 to 17 months).Cauda equina symptoms were observed in one patient and compression fracture of the adjacent vertebra happened in 2 patients,but no infection or pulmonary embolism occurred in any patient.Conclusions Both high viscosity cement PVP and low viscosity cement PVP can relieve back pain and improve quality of life in patients with severe OVCF,but the former may increase the efficacy and safety of PVP obviously in correction of cobb's angle,height restoration of the injured vertebra and reduction of cement leakage.

Purpose To investigate the expression and significance of androgen receptor (AR) in breast carcinoma with different ER and PR status.Methods Immunohistochemical method was used to detect AR, ER and PR expression in 173 cases of breast carcinoma, and all the cases were grouped according to: (1) AR status: AR~- positive subgroup and AR~- negative subgroup, (2) ER status: En subgroup (negative for both ER and PR), Ep subgroup (positive for ER and/or PR), (3)AR, ER and PR status: En-AR~+ subgroup (AR~- positive cases of En subgroup), En-AR~- subgroup (AR~-negative cases of En subgroup), Ep-AR~+ subgroup (AR~- positive cases of Ep subgroup), and Ep-AR~- subgroup (AR~-negative cases of Ep subgroup).En-AR~- subgroup also was called negativE-for-all subgroup, and the rest three subgroups were called partly-or-completely positive subgroup (positive for at least one of the three markers), and then, the groups were compared with clinicopathological features.Results The positive rate of AR was 62.8% (54/86) in Ep subgroup and 37.9%(33/87) in En subgroup, and there was significant difference between them (P=0.001).Compared to AR~-negative subgroup, AR~-positive subgroup had smaller tumor size, less mitosis count and lower grade(P<0.05).Compared to the other three subgroups, En-AR~- subgroup had more mitosis count and higher grade (P<0.01).In En subgroup,AR~- positive cases had less mitosis count and lower grade (P<0.05), and in Ep subgroup,AR~- positive cases had higher stage(P=0.000), but no significant difference were found among partly or completely positive subgroups.Conclusions The expression of AR may play a different role in breast carcinoma with different ER and PR status, because it indicates good prognosis in negative for both ER and PR subgroup, but bad in ER and/or PR positive subgroup. When choosing a personalized therapy, all the combinations of hormone receptor status should be considered for the patients.

OBJECTIVE: To establish an HPLC method for the determination of the contents of the related substances in metoclopramide.METHODS: The sample was separated on C18 column with the mobile phase consisted of acetonitrile-0.02 mol?L-1 phosphonic acid solution(19∶81) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 275 nm;the column temperature was kept under room temperature and the injected volume was 20 ?L.The contents of the related substances in metoclopramide were computed by self-control method of main constituent.RESULTS: Under the above described chromatographic conditions,metoclopramide was completely separated from its impurities.A good linearity between impurities' peak area and metoclopramide(contrast solution) concentration was achieved when the concentrations of impurities were over the range of 0.05%～4.0%.The lowest detectable limit of metoclopramide was 0.3 ng,and the contents of the related substances were all less than 0.26%.CONCLUSION: The method is convenient,accurate,sensitive and,specific,and it can be used for the determination of the related substances in metoclopramide.