To increase the biotransfomation efficiency from the orotic acid to the uridine 5'-monophosphate(UMP),URA5 gene encoding orotate phosphoribosytransferase was amplified from Saccharomyces cerevisiae BY4742 by PCR,then it was inserted into the expression vector pYX212(contained orotidine monophosphate decarboxylase gene URA3)and the pYX212-URA5 was transformed into Saccharomyces cerevisiae BJX12 by electroporation.The recombinant strain was elementarily used to convert orotic acid to UMP.The results showed that pYX212-URA5/BJX12 could accumulate 7mmol/L UMP from 32mmol/L orotic acid in 26h,significantly higher than both control groups pYX212/BJX12(2.7mmol/L) and BJX12(2.4 mmol/L).

Objective To analyze clinical features,surgical treatment efficacy and prognostic factors of bone metastasis patients after liver transplantation for hepatocellular carcinoma.Methods A retrospective clinical data of 20 bone metastasis patients after liver transplantation for hepatocellular carcinoma from July 2000 to January 2010 were received.The effect of surgery aimed at bone metastasis was evaluated.Univariate and multivariate prognostic risk factors were analyzed.Results The median survival time of these patients was 7.5 months and 1-year survival rate was only 20％.Surgical treatment could relieve pain and promote patients' peformance status significantly.Univariate and multivariate analysis found that tumor microvascular invasion within the removed recipient liver was the only prognostic risk factor.Conclusions Patients of bone metastasis after liver transplantation for HCC had poor prognosis.Surgical treatment helps improve patient's quality of life.Tumor microvascular invasion is the risk factor of surgical prognosis.

Objective To study the effect of Yuan‐source Points and Collateral‐points combination and Bushen Yishou cap‐sule on hemorheology and hs‐CRP in patients with cerebral infarction .Methods A total of 120 patients with neurological function defects diagnosed with cerebral brain infarction were randomly divided into 3 groups ,who were given general treatment and nursing as usual ,Acupuncture group in general treatment based on Yuan‐source Points and Collateral‐points combination for enjoy points principles ,to acupuncture treatment ;drug group to oral Bushen Yishou capsule;combined group to Yuan‐source Points and Collat‐eral‐points combination and oral Bushen Yishou Capsule .Neurological function defects scores ,blood rheological properties and blood hs‐CRP were evaluated in all patients before and after treatment for 1 month .Results Three sets of results in treatment of limb function after scoring ,hemorheology and serum hs‐CRP levels were significantly improved (P<0 .01) .There was better improve‐ment in Combined group than these in Acupuncture group and Drug group respectively (P<0 .01) .Conclusion Yuan‐source Points and Collateral‐points combination combined with Bushen Yishou capsule can significantly improve neurological function and hemo‐rheology ,hs‐CRP levels in patients with cerebral infarction .

Objective To observe the changes of serum calcium, phosphorus, calcium-phosphorus product and para-thyroid hormone(PTH)levels after treatment with different concentrations of calcium dialysate in combination with calcitri-ol. Methods Thirty-six patients on maintenance hemodialysis were randomly divided into Dca1.25 group and Dca1.5 group, 18 cases in each group. Patients took different doses of calcitriol on the different values of PTH basis. Changes of se-rum calcium, phosphorus, calcium-phosphorus product,PTH and bone alkaline phosphatase (BAP) levels were recorded re-spectively on four time points from prior treatment and 1, 3 and 6 months of therapy. The levels of systolic blood pressure (SBP) and diastolic blood pressure (DBP) were also observed. Results There was an interaction between treatment effects and time effects in serum calcium, phosphorus, calcium-phosphorus product levels in two groups (P＜0.05). With the dura-tion of treatment, there was a trended to decrease and increase respectively in DCa 1.25 group and DCa 1.5 group, and trend-ed to stabilize for therapy 3 to 6 months. There was no interaction between serum PTH and BAP levels and time effects in two groups. There was no statistical difference in simple treatment effects and time effects. The serum PTH level showed a trend of increase gradually in DCa1.5 group with the duration of treatment. In DCa 1.25 group, the serum PTH level trended to stabilize after 3-month therapy and showed a trend of decrease gradually. There was an interaction between treatment effects and time effects in SBP in two groups, but no interaction in DBP. There was a significant difference in SBP before and after treatment in two groups. The level of SBP reduced tardily with time of treatment and reduced significantly after 3 and 6-month therapy. Conclusion The application of DCa1.25 dialysate in combination with calcitriol decreased serum calci-um effectively and kept the lower limit of normal value, which increased tolerability of active vitamin D and calcium carbon-ate therapy, and decreased SBP in maintenance hemodialysis patients.

Objective To explore the risk factors and metastasis of non-sentinel lymph node (NSLN) in breast cancer patients with sentinel lymph node metastasis≤2. Methods 88 cases of breast cancer with sentinel lymph node metastasis≤2 were retrospectively analyzed. Factors influencing the status of NSLN were studied. Results 8 cases (9.09%) had NSLN metastasis. Variance analysis showed that tumor stage(P=0.014), histolog-ical grade(P=0.014) and ER statuts(P=0.009) were correlated with NSLN metastasis. Logistic analysis showed that only histological grade was the independent predictor for NSLN metastasis (P=0.016,OR=1.150,95%CI=0.598~7.740).18 cases had micro-metastasis of sentinel lymph node and 70 cases had macro-metastasis of sentinel lymph node. The ratio of false negative SLN was 10.23%. The mean follow-up time was 11 months. There was no case of recurrence, metastasis or death. Conclusions The metastasis rate of NSLN is relatively low in breast cancer patients with small number of positive SLNs. NSLN metastasis is related to tumor stage, histological grade, and ER expression.

ObjectiveTo investigate the effects of methylprednisolone （MP） on the proliferation and expression of glial fibrillary acidic protein (GFAP) in astrocytes after spinal cord injury in rats. MethodsThe spinal cords in T8～T10 in 48 male Wistar rats (10 weeks old) were exposed and the right site were semi-transected. 24 rats were administrated MP through tail vein as experiment group, and the other rats were injected equal volumes of normal saline as the control group. They were assessed with Basso, Beattie, and Bresnahan (BBB) score for ethological test every 4 d, and were sacrificed, perfused 3 d, 7 d, 14 d and 28 d after operation. The spinal cords were removed and postfixed, The expression of GFAP was observed with immunohistochemical staining and gray values determination. The GFAP positive cells were also counted. ResultsThere was no significant difference in BBB score between these groups until 21 d after operation; but the MP group rats showed significantly better functional recovery than the control group 25 d after operation. The number of positive astrocytes was less in the MP group than in the control group 3 and 7 d after operation, but there was no significant difference 14 d after operation. The GFAP expression in glial scar was significantly lower in the MP group than in the control in the first 21 d after operation, but no significant difference 28 d after operation. ConclusionThe large dose methylprednisolone administration can decrease the number of active AS, attenuate the formation of glial scars, and improve the functional recovery of the hind limbs after acute spinal cord injury.

Objective:To analyze the differentally expressed long non-coding RNA (lncRNA) among mice of different ages and explore the mechanism of kidney aging.Methods:Male C57BL/6 mice aged 3-month-old ( n=5), 12-month-old ( n=5) and 24-month-old ( n=5) (each weighting about 25 g) were randomly selected. PAS staining, Masson staining and senescence associated β-galactosidase (SA-β-gal) staining were used to detect the pathology and cell senescence of mice kidney. High throughput sequencing was performed to detect the differentially expressed lncRNA and their fragments per kilobase million. Real-time quantitative PCR was used to verify the differentially expressed lncRNA. Competitive endogenous RNA (ceRNA) network, which consisted of lncRNA, miRNA and mRNA was built. GO and KEGG enrichment analysis method were used to predict the biological function of differentially expressed lncRNA. Results:PAS staining and Masson staining showed the development of kidney fibrosis, and SA-β-gal staining positive region was increased significantly as age increased. There were 938 known lncRNA and 542 novel lncRNA differentially expressed among different ages' mouse kidney. Compared with 3-month-old mice, 33 lncRNA were up-regulated and 43 lncRNA were down-regulated in 12-month-old mice. Compared with 3-month-old mice, 130 lncRNA were up-regulated and 91 lncRNA were down-regulated in 24-month-old mice. Compared with 12-month-old mice, 36 lncRNA were up-regulated and 22 lncRNA were down-regulated in 24-month-old mice. The results of qRT-PCR about verified 10 lncRNAs with larger differential expression multiples and higer expression levels were consistent with the sequencing data. GO enrichment analysis showed that the target genes of lncRNA differentially expressed in the three groups were mostly located in the nucleus and cytoplasm, and might play a role by binding to proteins or participate in various protein phosphorylation, cell cycle, transcription, transcription regulation and other processes. KEGG enrichment analysis showed that the target genes of lncRNA differentially expressed in the three group were significantly enriched in Rap1 signaling pathway, FOXO signaling pathway and MAPK signaling pathway, which were closely related to kidney aging.Conclusion:There are significant differences in expression of lncRNA among the kidney of different ages mice, which are involved in the occurrence of renal senescence.

Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.

Objective To investigate the spatial distribution and clustering areas of Budd-Chiari syndrome in Heze City,Shandong Province,and to provide epidemiological information for further exploring the etiology and related risk factors of the disease.Methods Detailed residential addresses of 342 cases of patients (residents of Heze City) with diaphragm type Budd-Chiari syndrome diagnosed between 1995 and 2004 in Heze Municipal Hospital,Heze Shan County Central Hospital,Affiliated Hospital of Xuzhou Medical College,Shandong Provincial Hospital and Beijing Xuanwu Hospital were collected.Geographic information system (GIS) was used as a platform for data management and display.The nearest neighbor index,Ripley's K(d) function,Ripley's L(d) function and the nearest neighbor clustering method were applied to detect the spatial characters of Budd-Chiari syndrome in Heze City,Shandong Province.Crimestat 3.0 was used for spatial analysis.Results The nearest neighbor distance analysis showed that the nearest neighbor index was 0.6767 (Z =-11.4387,P ＜ 0.01).That was an aggregation at the first-order spatial scale.Within the study area,the first clustering radius of Budd-Chiari syndrome was 6.66 km,and the first clustering strength was 5.40; the average radius of the strongest clustering area was 126.61 km,and the clustering strength was 12.52,while the biggest clustering radius was larger than 222 km.After corrected by population,the gathering strength was slightly higher than that before the correction.Ten first-order hot spots were formed,and 95％ confidence interval aggregation number was 7,which meant the results were statistically significant(P ＜ 0.05),main clustering areas are in Mudan District,Shan County and Juancheng.One secondorder hot spot was gathered based on the first-order hot spot.Conclusions Spatial distribution of Budd-Chiari syndrome in Heze City,Shandong Province has showed spatial aggregation and heterogeneity.This study has a great epidemiological significance for further exploring the cause of Budd-Chiari syndrome.

Objective To evaluate the functional and oncological outcomes of patients with locally advanced prostate cancer (PCa) treated by hormone therapy combined with extra-fascia1 laparoscopic radical prostatectomy (LRP) plus extended lymph node dissection (ePLND).Methods From January 2004 to June 2016,a total of 255 PCa cases (pT3-4NxM0) who received LRP plus ePLND were enrolled into our study.The mean age of the patients was 67 (range 44-88) years,and median PSA level was 21.2 (range 0.6-454.0) ng/ml.The patients were divided into earlier group (from January 2004 to December 2011,160 cases) and later group (from January 2012 to June 2016,95 cases) according to different treatment periods.The baseline demographics between the two groups were similar.All patients routinely received adjuvant hormone therapy (AHT) postoperatively.The patients in the later group underwent collapsin response mediator protein 4 (CRMP4) methylation study on the prostatic biopsy preoperatively.Those with a CRMP4 methylation level ＞ 15％ or rectum/bladder neck invasion,were treated by neoadjuvant hormone therapy (NHT) for 3-6 months.Positive surgical margin (PSM),progression-free survival (PFS),cancer-specific survival (CSS),overall survival (OS) and postoperative continence rates between the two groups were analyzed and compared.Results The mean operative time of the earlier and later group were (239 ±65) min and (203±51) min,mean blood loss were (109-±65) ml and (96-±44) ml,mean dissected nodes were (19 ± 5) and (21 ± 7),respectively (all P ＞ 0.05).The total PSM rate was 19.2％,and PSM rates of the two groups were 23.1％ and 12.6％ (P =0.04).All the 255 cases received AHT and 25 cases in the later group underwent NHT.The median follow-up time was 73 months (range 10-152 months).The total 5-year PFS,CSS and OS rates were 77.7％,94.3％ and 87.1％,respectively,and the rates between groups were 73.8％ vs.86.1％ (P=0.03),93.1％ vs.98.6％ (P=0.07),and 85.0％ vs.92.8％ (P =0.11),respectively.The 1-year postoperative continence rates were 91.9％ vs.97.9％ (P =0.09).Conclusions Hormone therapy combined with LRP plus ePLND represents an oncological and functional effective option in patients with locally advanced PCa,and improved PFS might be acquired by preoperative tumor staging.