Objective To construct a cDNA subtractive library of genes transactivated by c-terminally truncated middle surface protein of hepatitis B virus(MHBs t) with suppression subtractive hybridization technique for cloning genes associated with transactivation. Methods The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-Mt167 and pcDNA3.1(-) empty vectors, respectively, then cDNA was synthesized. After restriction enzyme Rsa I digestion, small-size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results The subtractive library of genes transactivated by MHBs t was constructed successfully. The amplified library contained 94 positive clones. Colony PCR showed that these clones contained 200-800bp inserts. Sequence analysis was performed in 50 clones,and the full length sequences were obtained with bioinformatics method. 23 coding sequences were obtained in total, which consisted of 19 known and 4 unknown ones.Conclusions The obtained sequences may be target genes transactivated by MHBs t, among which some genes coding proteins may involve in cell cycle regulation, immune response and tumour genesis.

Objective To study the effect of combination mode of blood purification on insulin resistance in MODS patients and discuss the best mode of blood purification to improve insulin resistance in MODS patients.Methods A total of 60 MODS pa-tients were selected and randomly divided into control group(single mode of blood purification)and treatment group(combination mode of blood purification),each group of 30 cases.Blood glucose (BG),glucose standard deviation (BGSD),coefficient of variation of blood glucose (BGCV),regular insulin dosage (Ins),fasting insulin (FINS),insulin resistance index (HOMA-IR),HbA1c,C-re-active protein (CRP)was observed before and after treatment.Results BG,HbA1c,FINS,CRP,HOMA-IR,Ins of experimental group was lower than that of control group (P <0.05).After treatment,BG,FINS,CRP,HOMA-IR,Ins was declined in experi-mental group (P <0.05).After treatment,HbA1c was not changed in experimental group (P >0.05 ).Conclusion Combination mode blood purification may be more effective to insulin resistance in MODS patients.

Objective To screen stage\|specific expression genes of plerocercoid of Spirometra erinacei\|europaei. Methods RNA was extracted by the acid guanidinium thiocyanate\|phenol\|chloroform from plerocercoid and adult worm of Spirometra erinacei\|europaei. Contaminated DNA in the RNA was digested by RNase\|free DNase. cDNA was synthesized by using T\-\{12\}MA, T\-\{12\}MC, T\-\{12\}MG and T\-\{12\}MT primers, and PCR was then done using the same T\-\{12\}MN and one random primers. The PCR products were fractionated on 8% denatured polyacrylamide gel, differential bands of plerocercoid found in the gel were cut out, amplified by PCR and sequenced after the gel was dried out and autoradiographed. Northern hybridization was conducted to identify the stage\|specific expression genes. Results Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization after they were amplified by PCR. The fragments 1 and 2 were confirmed to express specifically in plerocercoid by Northern hybridization, but the fragment 3 was expressed in both plerocercoid and adult worm. When the 3 gene fragments were homologically analyzed in GenBank, the sequence which was homologous with the fragments 1 and 2 was not found, but the fragment 3 had high homology with many kinds of 28S rRNA. Conclusion The gene expression of plerocercoid was different from that of adult worm probably because they live in different hosts. Two kinds of different gene fragments in plerocercoid were identified by mRNA differential display technique.

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, which provides a new therapeutic idea for myocardial injury.OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P <0.05) and its membrane potential decreased significantly compared with the blank control group (P < 0.05). Theapoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantlycompared with the 5-azacytidine group (P < 0.05, P < 0.01), and their membrane potential increased significantly (P <0.05, P < 0.01). The results of western blot showed that the expression level of Nkx2.5 increased significantly in thepilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while theexpression levels of GATA4 and ATF-2 in each group were low and there were no significant differences among groups(P > 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4,ATF-2 and MEF-2C needs to be further studied.

Allicin,which contains rich sulfide ,is provided for prevention and treatment of tumor ,anti in-flammation,prevention and treatment of cardiovascular diseases ,improving immunity and so on .Studies show that allicin can not only prevent many types of cancer ,but also has obvious inhibitory effect on tumor .The mechanism is related with antioxidant effects ,reducing the invasion of tumor cells ,inhibiting the proliferation of tumor cells , inducing tumor cell apoptosis and directly killing the tumor cells .This paper reviews the research progresses on antitumor allicin .

Objective To investigate the status of nursing professional values and its influencing factors to make contribution for nursing managers to improve nursing professional values.Methods 280 registered nurses selected by convenient sampling method were investigated by demographic questionnaires and the Revised Nursing Professional Values Scale (NPVS-R).Results The score of nursing professional value was (100.70 ± 6.36) points,the highest scoring dimension wasprofessional characteristic ,followed by trust, behavioristand providing care.The scores of protecting patients privacy,being trusted by patients,constantly improving professional ability,etc.were higher.The scores of involving in research,peer review and public decision,etc.were lower.Medical department,profession and interest,job satisfaction were the influencing factors,F values were 2.575,3.750 and 2.909.Conclusions The nursing professional value was higher than middle level and unbalanced development.Nursing managers should take individualized approach to improve nursing professional values based on the characteristics of departments and lower score parts.

Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group NSCs group > control group, and between the groups was significant difference ( P < 0. 05 ) . Conclusions By Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene (bcl?2) gene transfection make neural stem cells can promote cultured rat neural stem cells. bcl?2 gene?modified neural stem cell transplantation can promote the regeneration of spinal cord injury synaptic elevated HSP27 expression after spinal cord injury, reduced expression and neural cell apoptosis after spinal cord injury bcl?2 gene and improve limb movement in rats function and electrophysiological function.

Objective To study the role of B-mode ultrasound found bladder trabeculation in evaluating the degree of bladder outlet obstruction (BOO) and the bladder function in benign prostatic hyperplasia (BPH) patients.Methods Conducted prospective research to determine differences in clinical data and urodynamic parameters between BPH patients with and without bladder trabeculation diagnosed by abdominal ultrasound.Results Thirty-six BPH patients with bladder trabeculation were compared with 68 BPH patients without bladder trabeculation.The mean age was (73.7 ± 10.1 ) years for the patients with bladder trabeculation and ( 69.6 ± 6.2 ) years without bladder trabeculation, IPSS was 24.4 ± 6.6 and 22.8 ± 8.3 respectively, in which no significant differences were found ( P ＞ 0.05 ).The detrusor pressure at maximum flow rate was ( 131.7 ± 57.3 ) cm H20 and (92.1 ± 47.8 )cm H2O ) respectively.The linearized passive urethral resistance relation was 4.6 :± 1.1 and 3.5 ± 1.5 respectively, showing a significant difference ( P ＜.05 ) between the two groups.Seventy-two point two percent (26/36) of the patients with bladder trabeculation had a low compliance bladder, among whom 23.1% (6/26) of the patients had bilateral hydronephrosis with renal insufficiency.The percentage for the control group was 42.6% (29/68) and 10.3% (3/29) respectively (P ＜ 0.05).Conclusions The bladder trabeculation found by B-ultrasound in BPH patients suggests the existence of BOO and a high risk of bilateral hydronephrosis.Bladder trabeculation in patients without urinary retention suggests they are in compensation status.Relief of the obstruction is helpful to recovery of bladder function and the reduction of complications.

Objective To investigate the efficacy and safety of low dose of oral desmopressin in elderly women with nocturia. Methods Eligible female patients with nocturia older than 60 years were included in this study.A total of 97 patients were randomly divided into 2 groups.Care was taken to match the patients of the two groups by age and clinical criteria.Control group (n=48 ) received liquid restriction during nighttime.Experimental group (n=49) received 0.1 mg desmopressin at bedtime and liquid restriction for 8 weeks.Patients were required to visit the outpatient clinic from the first visit,and after 4 and 8 weeks of treatment.Patients maintained flow volume charts and used diaries to record voiding data throughout the study.At each visit,all patients were evaluated by blood biochemical routine test,mean nocturnal urine volume,mean number of nocturia,mean duration of the first sleep period and sleep quality.At baseline,all the patient's blood sodium,liver and kidney function were normal. Results After 4 weeks of treatment with desmopressin,28 patients (57.1％) had less than 2 voids.15 patients (31.3％) in the control group had less than 2 voids.After 8 weeks,35 patients (71.4％) with desmopressin had less than 2 voids.16 patients (33.3％) in the control group had less than 2 voids.Compared with control group,nocturia cure rate in experimental group was significantly higher after 4 weeks and 8 weeks (P ＜ 0.05).After 8weeks,desmopressin significantly decreased mean nocturnal urine output from 590 ± 70 ml to 376 ± 50 ml (P ＜ 0.05).Mean nocturnal urine output in the control group was not significantly decreased from 600 ± 90 ml to 550 ± 60ml (P ＞0.05) ; Mean number of nocturia before and after receiving desmopressin were 2.9 and 1.6 respectively which differed significantly (P ＜ 0.05).Mean number of nocturia before and after in control group were 2.8 and 2.3 respectively with no significant difference (P ＞ 0.05).The mean duration of the first sleep period increased by 73％ (from 2.2 to 3.8 h) in the desmopressin group,compared with an increase of 19％ (from 2.1 to 2.5 h) in the control group (P ＜ 0.05).39 (79.6％) patients in demopressin group were satisfied with sleep quality compared with only 15 (31.3％) patients in control group were satisfied (P ＜ 0.05).No serious complications were found during the medication.All the patient's blood sodium,liver and kidney function remained normal during treatment. Conclusions Low does oral administration of desmopressin could be an effective and well-tolerated treatment for nocturnal polyuria in elderly women.

Objective To analyze the risk factors of restenosis after renal artery endovascular treatment in patients with Takayasu arteritis.Methods In this study,39 patients with Takayasu arteritis underwent endovascular therapy from January 2003 to March 2014.Univariate and multivariate logistic regression analysis were used to analyze the risk factors relating to restenosis.Results There were 13 males and 26 females.The mean age was (27 ± 11) years.Seventeen unilateral and 22 bilateral renal artery stenosis were treated.A total of 54 endovascular procedures were successfully performed including 23 cases of unilateral percutaneous transluminal angioplasty (PTA),14 cases of unilateral stent implantation,12 cases of bilateral PTA,and 5 cases of bilateral stent implantation.The mean follow-up was (48 ±34) months,the restenosis rate was 38.9％ (21.54) and the mean time of restenosis was (11.5 ± 2.3) months.Elevated erythrocyte sedimentation rate (ESR) (OR =6.624,95 ％ CI:1.222-35.902) was independent risk factors for restenosis.Antiplatelet therapy (OR =0.158,95％ CI:0.028-0.887) and glucocorticoids or i mmunosuppressive therapy (OR =0.035,95 ％ CI:0.003-0.349) were protection factors against renal artery restenosis.Conclusion The elevated ESR increases the risk of restenosis after endovascular treatment in Takayasu arteritis associated renal artery stenosis.Antiplatelet therapy and glucocorticoids or immunosuppressive therapy were protection factors for renal artery restenosis.