AIM: To explore the effect and the mechanism of sulphated heparin on the proliferation and the apoptosis of human hepatocellular carcinoma cells. METHODS: The human hepatocellular carcinoma cell line (HepG-2) was used to identify the expression of ras gene protein and to study the effect of sulphated heparin on proliferation and the apoptosis in vitro . RESULTS: The sulphated heparin downregulated the ras protein expression and inhibited the cell growth in HepG2 cells. In the presence of sulphated heparin, the apoptosis rate of HepG2 increased. CONCLUSION:The data suggest that the effects of sulphated heparin on the proliferation and the apoptosis of the human hepatocellular carcinoma cell are correlated with the signaling transduction mediated by ras gene protein.

AIM To study the direct effect of amiloride on recombinant human protein kinase CK2 holoenzyme and its kinetics. METHODS Recombinant human protein kinase CK2 ? and ? subunits were cloned,expressed and purified by gene engineering. The two subunits were mixed at the same molar ratio to form CK2 holoenzyme,which displayed the maximum biological activity. CK2 activity was assayed by detecting incorporation of 32 P of [? 32 P]GTP into the substrate in various conditions. RESULTS Amiloride inhibited the holoenzyme activity of recombinant human protein kinase CK2 with an IC 50 of 257 57 ?mol?L -1 . Kinetic studies of amiloride on recombinant human CK2 showed that the inhibition was competitive with GTP with the K i of 236 30 ?mol?L -1 ,and noncompetitive with casein with the K i of 163 63 ?mol?L -1 . CONCLUSION Amiloride inhibits the activity of cAMP or ATP dependent enzymes as well as the kinase with GTP as phosphate donor,and the recombinant human protein kinase CK2 may be used as a molecular target for simple screening and development of effective inhibitors of CK2.

Objective To summarize the experience in transanal local excision (LE) for rectal cancer (RC). Methods The clinical data of 28 cases of RC treated by LE from 1988 to 1998 were analyzed retrospectively. Results In this series, five-year survival rate was 83.4?6.2%, and the local recurrence rate (LRR) was 17.8%. In well-differentiated carcinoma, 4 cases were convinced as local recurrence with a LRR of 17.4%(4/23); in moderately- differentiated carcinoma, one case with a LRR of 20.0%(1/5). The LRR in T 1 and T 2 group was 15.0% (3/20) and 25.0% (2/8) respectively. LRR was 16.7% (4/24) in patients with less than 1/3 bowel wall involved, LRR was 16.7%(4/24),whereas LRR was 25.0%(1/4) in more than 1/3 bowel wall involved group. In total bowel wall resection group the LRR was 16.7%(3/18) while in partial resection group was 20.0% (2/10). In patients with tumour size larger than 4 cm LRR was 22.2% (2/9), tumour size smaller than 4 cm LRR was 15.7% (3/19). Conclusion LE for RC might only be successfully performed in selected patients (T 1～T 2, N 0M 0, well or moderately-differentiated carcinoma,low RC within 6 cm from anal edge). The indications of transanal LE must be controlled strictly. Total excision of tumor and prevention of implantation of carcinoma are the main points in the prophylaxes of recurrence. Postoperative follow-up is needed in order to find local recurrence as early as possible.

Objective To investigate the expression of autophagy and the effect of complement C5a/C5aR pathway on autophagy induced by renal ischemia reperfusion injury (IRI).Methods MaleWT and C5aR gene knockout (BALB/C background) mice were selected.The model of renal IRI was established by occluding bilateral renal pedicles with microaneurysm clamps.Mice were divided into wild type BALB/C (WT) group and C5aR gene knock out (C5aRKO) group.The pathology of kidney was assessed by HE staining.The levels of BUN and KIM-1 were detected 24 h after reperfusion.The expression of the autophagy-associated protein (LC3 Ⅱ/LC3 Ⅰ and P62) was measured by Western blotting and immunofluorescence.In vitro,human renal tubular epithelial cells (HK2) were cultured.The expression of LC3 in HK2 cells was investigated by immunofluorescence and Western blotting after being treated with recombinant C5a or C5a combined with C5aR antagonist (C5aRA).Results As compared with WT group,the severity of kidney injury was obviously reduced in C5aRKO group (P＜0.05).After ischemia-reperfusion,the expression of autophagy-related protein LC3 gradually increased with the reperfusion time prolonged.The level of autophagy induced by ischemia-reperfusion was significantly reduced in C5aRKO group as compared with WT group (P＜0.05).In addition,the expression of autophagy-related protein LC3 Ⅱ in HK2 cells was increased with the augment of C5a stimulation concentration in vitro.Blockage of C5aR pathway by C5aRA led to a significant decrease in autophagy (P ＜ 0.05).Conclusion Complement C5a/C5aR pathway promotes renal IRI-induced autophagy.

Objective To understand the influencing factors of professional adaptability for college nursing students and the mechanism. Methods A group of 1 105 college students from grades 2010 and 2011 were surveyed with questionnaire for professional adaptability. The data were analyzed with SPSS 16.0. Measurement data were expressed as x±s while enumeration data were expressed as percentage. t test and variance analysis were used to compare the differces among groups of different grades and genders. Results The professional adaptability mean score of college nursing students was (102.66 ± 14.93) and significant differences in professional adaptability were observed between different grades and genders (P=0.004 3, P=0.001 5). The possible influencing factors professional adaptability were different sources of students, family location, family income, etc. There were statis-tic differences in professional promise between arts and science students and between rural an urban students (P=0.029 8, P=0.007 0). Conclusion Grade and genders are the main influencing factors for professional adaptability of college nursing students while other factors exert little effect on it.

Objective To investigate the expression of proinflammatory cytokine interleukin-17(IL-17) and complement cleavage fragment C5a and the regulation effect of C5a on the expression of IL-17 during renal allograft rejection .Methods The frequency of IL-17+ T cell in peripheral blood and the expression of IL-17 in renal tubular epithelial cells (HK2) after C5a stimulation in renal transplant recipients were measured by flow cytometry and the changes of serum C5a level was detected by ELISA ,respectively .Im-munohistochemistry was applied to detect and compare the expression of IL-17 and the deposition of C5b-9 in normal renal tissues and renal tissues with allograft rejection .The difference of IL-17 expression in HK2 cells before and after the recombinant C5a stimulation was detected by immunocytochemistry .Results Both the percentage of IL-17+ T cells and serum C5a levels were sig-nificantly increased after the allogeneic renal transplantation .Compared with normal renal tissues ,both the deposition of C5b-9 and the IL-17 expression in renal tissues with allograft rejection was remarkably up-regulated ,which showed the positive correlation be-tween them .The expression of IL-17 in HK2 was obviously up-regulated by the recombinant C5a stimulation .Conclusion C5a may positively regulate the expression of IL-17 by tubular epithelial cells during the renal allograft rejection .

Objective To evaluate the graduation clinical skill examination based on objective structured clinical examination(OSCE) for college nursing undergraduates.Methods Classic OSCE was simplified and taken as references for nursing undergraduate graduation clinical skill examination including nursing evaluation,nursing dignosis,nursing intervention and health education for stations.Self-made questionnaire was conducted among students.Examination scores and the questionnaire results were statistically analyzed by SPSS 16.0 software.Measurement data and categorical data were shown as ((x)±s) and ％ respectively.T test was used in comparison between the groups according to the inspection level of α =0.05,P ＜ 0.05 means significant difference.Results Simplified OSCE was composed of 4 stations and the mean score was (80.38 ± 4.22) with mean scoring rate of the 4 stations varied from 72.2％ to 88.0％.More than 93％ students thought that OSCE score can or basically can evaluate comprehensive clinical nursing competence,clinical thinking,communicating and adaptability.Conclusions Simplified OSCE is designed properly for the objective evaluation of nursing students' clinical competence and students are mostly satisfied with its design and organization,quality and efficiency.

Objective To measure the costs of primary health services in the community for developing government compensation standard and prediction of budget in 201 5.Methods Literature review and panel discussions were used to build the index database.A two-round Delphi expert consultation determined the work to do and steps for community healthcare standards.60 community health centers were sampled by stratified random sampling for survey,and the work hours,workload and service volume of respective services in 2013 were measured according to the service standards of primary care.Results The community primary care consists of the categories of outpatient services,nursing care, laboratory tests and drug management,totaling 20 services and 88 working procedures.The total work hours of primary healthcare service at 60 community health centers were 2 557 187.9 hours,which is adjusted to 2 959 21 5.1 hours based on workload coefficients.Based on the income standards of employees at the community health centers investigated,and the human cost price which was recommended by the experts,the total costs of primary healthcare of the 60 centers were 245 million,248 million and 318 million respectively,and the average cost was 71.0 yuan,71.8 yuan and 92.1 yuan per visit respectively.By such standards,the government should subsidize 38.9 ~ 60.0 yuan per visit to the centers.The total expenditure of primary healthcare of all communities in Shenzhen in 201 5 was predicted to be 314 to 407 million,for which the government is expected to subsidize 1.68~2.58 billion.Conclusion The subsidies for community primary healthcare fall short as the cost per visit runs up in 2013 to 71.0 yuan per visit or more,far above the current subsidy of 32.1 yuan per person.The subsidy per visit should be made 50.0 yuan in 201 5.

OBJECTIVETo investigate the insulin-like growth factor-I (IGF-I) gene and polypeptide expression in cultured rat osteoblast (ROB) and the role of IGF-I in mediating the cell-to-cell communication by mimicking the pharmacokinetics of parathyroid hormone (PTH).

METHODSThe ROB was cultured with three kinds of treatment: (1) Control (Ctr), the cells were cultured without PTH during the first 6 hours and the subsequent 42 hours in a 48-hour cycle; (2) Intermittent exposure to PTH (Itm), the cells were cultured with PTH during the first 6 hours, but without PTH in the subsequent 42 hours; and (3) Continuous exposure to PTH (Ctu), the cells were cultured with PTH during the first 6 hours and the subsequent 42 hours.

RESULTSThe bone-forming activities of ROB were increased in Itm and inhibited in Ctu. The IGF-I mRNA content in Itm cells was elevated only during the first 6 hours and that in Ctu cells was elevated at any time during an incubation cycle. The free IGF-I concentration in the medium of Itm cells was generally higher and that of the Ctu cells was generally lower compared with those of the Ctr cells. The IGF-I antibody significantly reduced the alkaline phosphatase activity within the cells of Ctr and Itm.

CONCLUSIONSPTH rapidly and constantly stimulates the IGF-I gene transcription of osteoblast. There was an obvious discrepancy between the IGF-I mRNA content within the osteoblast and the free IGF-I level around the osteoblast in either mode of PTH action. The IGF-I might be important for osteoblast-osteoblast communication and bone-forming activity, not only in intermittent PTH administration, but also in the physiological functioning of osteoblasts.

Animals ; Cells, Cultured ; Gene Expression ; drug effects ; Insulin-Like Growth Factor I ; genetics ; physiology ; Osteoblasts ; Parathyroid Hormone ; pharmacology ; Peptides ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transcription, Genetic ; drug effects

Objective To study the validity of RAND-UCLA (Rand Corporation and University of California at Los Angeles) consensus panel method in developing guidelines of referral indications for low back pain (LBP).Methods Evidence-based clinical guidelines for LBP management at community level and its referral guidelines published since 2001 and other tools were retrieved with varied tools.All clinical guidelines met inclusion criteria were evaluated with clinical studies and evaluation tools (AGREE).An pool of indication items was established based on evidence for developing referral indications for LBP, which were added by RAND-UCLA consensus panel method, and alternative referral indications were selected and clinical guidelines for LBP referral were established.Results A total of 15 copies of clinical guidelines from nine countries or regions were included in it after critical appraisal.Four copies of referral guidelines from two countries were included.Referral indications for LBP were derived directly from the RAND-UCLA consensus panel process, consisting of 44 referral indications for three groups (immediate, urgent and routine referral).Conclusions The RAND-UCLA consensus panel method is a more useful and practical tool in developing clinical guidelines, referral guidelines, which is worthwhile being recommended and spread.