Object To optimize the apheresis procedure of mononuclear cells from bone marrow for different clinical treatments. Methods Bone marrow mononuclear cells were separated from bone marrow using COBE Spectra apheresis system. Cell morphology, cell counts, positive expression of CD34, CD133 and CD271 were detected, and the obtaining percent of total nuclear cells and mononuclear cells were calculated. The relationships between cell-morphology and CD34, CD133 and CD271, were also analyzed. Results 1)The positive expression of CD271 was correlated with monocytes before and after apheresis (P

Objective To evaluate the effects of smecta in prolonged hemorrhagic shock.Methods The modified Wigger's method was used to induce hemorrhagic shock in anesthetized rabbits.Twenty-nine rabbits were randomly divided into hemorrhagic shock group(n=14),smecta group(n=15),with smecta solution being administered via a gavage tube before shock.The plasma levels of endotoxin,tumor necrosis factor ?(TNF?),interleukin-6(IL-6) and nitric oxide(NO) were detected at pre-shock and post-shock,immediately after resuscitation and 2h after resuscitation.Blood culture was done at pre-shock,immediately after resuscitation,2h after resuscitation.The survival rates of 24h and 48h were observed.Results The plasma levels of endotoxin,TNF?,IL-6 and NO markedly increased after shock,and were maintained at high level in shock group (P

Objective To evaluate the effects of aminoguandine(AG) used as a selective inhibitor of the inducible isoform of nitric oxide synthase(iNOS) and L NAME used as a non selective inhibitor of nitric oxide synthase(NOS) on severe hemorrhagic shock Methods Forty rabbits were bled to mean arterial pressure of 4 4 66 kPa via femoral artery Hypotention was maintained for 120 min, the shed blood was then returned, followed by an infusion with Ringer′s solutions Animals were randomly divided into three groups: hemorrhagic shock group (n=14), AG group (AG 20mg/kg was infused intravenously ,n=14), L NAME group (L NAME 30mg/kg was infused intravenously with ,n=12) Plasma levels of endotoxin, tumor necrosis factor(TNF?) and nitric oxide (NO) were determined before and after shock, immediately after resuscitation and 0 5h, 2h, 4h after resuscitation The 24h and 48h survival rates were recorded Lung, intestine, liver and kidney tissues were obtained 48 h after shock for microscopic examination Results The plasma endotoxin, TNF? and NO levels markedly increased and were kept at high levels after shock Lower plasma levels of endotoxin TNF? and NO, less tissue damages and high survival rates were observed in AG group as compared with those in L NAME group and shock group Conclusions The endotoxin,TNF? and NO play an important role in the development of irreversible shock AG is beneficial in the treatment of hemorrhagic shock, while L NAME does not improve the outcome of shock

ProTide technology is a kind of prodrug design strategy invented by the team of Christopher McGuigan. ProTides are aryloxyphosphoramidates (or aryloxyphosphonamidates) which contain a phosphorus atom combined with an amino acid ester and an aryloxy group. These prodrugs can efficiently cross the cell membrane and escape from the first rate-limiting step of phosphorylation, which afford effective solutions to the drawbacks of current nucleoside analogues. At present, ProTide technology has been extensively applied in the field of antiviral research. It has been successful in providing a number of approved drugs and clinical candidates, such as sofosbuvir and so much more, highlighting the promising future in drug discovery. This review summarizes the brief history and characteristics of ProTide technology, as well as its application in the exploration of antiviral drugs.

Objective To evaluate the causes,prevention and treatment ot laparoscopic comphca- tions in urologic surgery.Methods From May 2000 to May 2004,135 urologic laparoscopies were per- formed,including 17 transperitoneal laparoscopies and 118 retroperitoneal laparoscopies.Of the135 cases,26 had adrenal adenoma extirpation;31 had roof-removal and decompression of renal cyst;25 had radical ne- phrectomy;6 had pyeloplasty;2 had partial resection of renal capsula and encapsulation of the great omen- tum;2 had partial nephrectomy;4 had pelviureteral resection;13 had ligation of renal pedicle lymphatic ves- sel;2 had ureterolysis;7 had ureterolithotomy;12 had ligation of spermophlebectasis;5 had exploration,re- duction and fixation of undescended testis.The intra-and post-operative complications were retrospectively reviewed and clinically analyzed.Results Overall,14 cases(10.4%)had complications.Intraoperative complications occurred in 10 cases,including 6 cases of peritoneal injuries,which were clamped during oper- ation;4 of vascular injuries,which resulted in conversion to open surgery.Postoperative complications oc- curred in 4 cases,including 2 of subcutaneous emphysema,which was spontaneously absorbed at 5 and 7 d after operation;1 of incision seepage,which was drained for 40 d and then disappeared;and 1 of urine reten- tion,for which urethral catheterization was kept for 3 d and then the patient had voluntary voiding.No death occurred in this series.Conclusions Understanding the characteristics of laparoscopic complications in u- rologic surgery and systematic training of the surgeons can reduce the occurrence of complications.

Red-based recombineering has been widely used in Escherichia coli genome modification through electroporating PCR fragments into electrocompetent cells to replace target sequences. Some mutations in the PCR fragments may be brought into the homologous regions near the target. To solve this problem in markeless gene deletion we developed a novel method characterized with two-step recombination and a donor plasmid. First, generated by PCR a linear DNA cassette which comprises a I-Sec I site-containing marker gene and homologous arms was electroporated into cells for marker-substitution deletion of the target sequence. Second, after a donor plasmid carrying the I-Sec I site-containing fusion homologous arm was chemically transformed into the marker-containing cells, the fusion arms and the marker was simultaneously cleaved by I-Sec I endonuclease and the marker-free deletion was stimulated by double-strand break-mediated intermolecular recombination. Eleven nonessential regions in E. coli DH1 genome were sequentially deleted by our method, resulting in a 10.59% reduced genome size. These precise deletions were also verified by PCR sequencing and genome resequencing. Though no change in the growth rate on the minimal medium, we found the genome-reduced strains have some alteration in the acid resistance and for the synthesis of lycopene.
Chromosomes, Bacterial ; genetics ; DNA ; Endonucleases ; metabolism ; Escherichia coli ; genetics ; Genetic Engineering ; methods ; Recombination, Genetic ; Sequence Deletion

and persistence of the SIRS,retrieve the unbalance of the pro-/anti-inflammatory,and improve the severe disease conditions.Therefore,it provides a new and feasible way to reglate SIRS in the early phase of SAP.

Objective To investigate neural correlates of the generation,execution and inhibition of motor intention. Methods Behavioral performance and event-related potentials (ERP) of 14 participants were recorded when they performed a two-staged Go-Nogo task. At the stage of intention preparation,three kinds of the "what" cues were presented separately to the participants : (1) to press a response key by the left or the right hand (the instructed) ,(2) to decide freely which hand will be used (the free) ,(3) to wait for the second cue (the control). At the stage of intention execution,the "whether" cue suggested the participants execute (Go) or withdraw (Nogo) the previous intention either voluntarily or forcedly. Results The interaction effect between the "what" and the "whether" was significant(F(2,26) = 8.262, P=0.002). The forced responses were faster than those of voluntary action under both the instructed condition ((468.52 ±105.87)ms vs (546.67 ±146.33)ms) and the control condition ((536.78 ± 83.28) ms vs (589.75 ± 140.80) ms). No significant difference was found for the free condition((538.71 ± 127. 39) ms vs (561. 44 ± 146. 51) ms). At the stage of intention preparation,significant ERP effects between the instructed intention and the freely intention were revealed in bilateral frontal regions (150～200 ms) , frontoparietal area (300～700 ms)and right temporal area. At the stage of intention execution,ERP effects between voluntary inhibition and voluntary action were observed in left prefrontal area (160～220 ms) ,fro-nto-cento-parietal area and right frontoparietal area. (300～550 ms). Conclusion The motor intention may be generated in the prefrontal area and maintained in the frontoparietal network. The left prefrontal and fronto-centro-parietal areas inhibit intention in the early stage and the right frontoparietal area involves response inhibition lately.

Objective To investigate neural correlates of the retrieval of temporal order.Methods ERP of 19 channels was recorded from 17 healthy young students while they performed a recency judgment task,a recognition task and a visual discrimination task separately,which were based on Sternberg paradigm.One-way ANOVA of repeated-measurement and post-hoc paired comparisons were carried out for both the task factor(temporal order,recognition and control)and the temporal distance factor correspondingly.The latter contains 3 intervals (1,2 and 3)between the targets.Results The temporal distance effect was significant:the closer targets correspond to longer reaction time((745.5±106.1)ms,(711.7±102.2)ms,(653.7±81.5)ms;F_((1.28,20.48))=43.37,P=0.00]and higher error rate((62.8±7.0)%,(72.5±5.4)%,(84.5±6.4)%;F_((1.32,21.11))=135.99,P=0.00].Significant behavioral effect of the task factor was also found for both reaction time(F_((1.32,21.11))=135.99,P=0.00)and error rate(F_((2,32))=200.31,P=0.00).No significant behavioral difference was found between the temporal order group and the recognition group.The spatiotemporal pattern of statistical parametric mapping(SPM)suggested the ERP effects of temporal order were presented in left temporal region (260～270 ms),bilateral parietal-occipital areas(280～290 ms),and bilateral parietal lobes(300～400 ms)prominently.No ERP effect was found between the temporal order task and the recognition task.Conclusions The neural correlates of the temporal distance effect exist in bilateral parietal regions.Temporal order as well as object information processing involves similar cerebral regions during the retrieval period.

Objective To evaluate the interstital MR lymphography using positive lymphotropic contrast agent for differentiation of metastatic lymph nodes from inflammatory lymph nodes hyperplasm.Methods Eighteen New Zealand white rabbits weighted at 2.0-2.5 kg were used.Inflammatory hyperplastic popliteal lymph node model was established in 9 rabbits by injection of complete freund adjuvant into the interdigitial skin of the dorsal aspect of one hind leg,and tumor-bearing popliteal lymph node model was established in another 9 rabbits by injection of VX~2 tumor cell suspension.The popliteal lymph nodes of another hind leg of all 18 rabbits were assigned to the normal contral group.In each group,every rabbit underwent MR lymphography examination before and after the inoculation.Volumes of 0.2 ml of Gd[DTPA-bis(2-aminoethoxy)ethane]polymeric contrast agent(Gd-poly-DTPA-EOEA)injection were injected subcutaneously into the dorsal feet of both hind legs of two groups of rabbits.T_1-weighted 3D gradient-echo images were obtained,and source images were used to reconstruct images of MIP before and after the administration of agent.The maximum short-axis diameter(MSAD)of each popliteal lymph node was measured on the enhanced 3D MIP images,and the signal-to-noise ratio(SNR)measurement was performed in the largest popliteal node of each rabbit at each time point in delayed scan.Independentsamples t test was used to compare the sizes of popliteal nodes in MSADs between inflammatorily hyperplastic and tumor-bearing nodes after the inoculation,and the values of SNRs of popliteal nodes at each time point between inflammatorily hyperplastic,tumor-bearing and normal popliteal lymph nodes.Imaging results of the popliteal nodes were analyzed and correlated with pathological findings.Results All of the rabbits were successfully inoculated except of the 2 rabbits in tumor-bearing nodal group.The size in MSAD of 13 inflammatorily hyperplastic and 11 tumor-bearing nodes was(1.32±0.14)and(1.33±0.12)cm,respectively.There was no significant statistical difference between the sizes of the two groups(t=0.186,P＞0.05).At the time of 5,15,30,60,90,120 minutes after the injection of the agent,the value of SNR of 9 inflammatorily hyperplastic and 9 contralateral normal nodes was 17.31±0.37 and 17.19±0.29,27.42±0.50 and 27.39±0.48,38.44±0.47 and 38.19±0.27,37.10±0.09 and 36.97±0.10,36.32±0.61 and 36.20±0.80,34.60±0.44 and 34.71±0.32,respectively.There was no significant statistical difference between the values of the two groups(t=0.78,0.14,1.43,1.00,0.36,-0.62,respectively,P＞0.05).The value of SNR of seven tumor-bearing and seven contralateral normal nodes was 6.00±0.22 and 17.21±0.28,7.87±0.16 and 27.17±0.23,8.14±0.24 and 38.16±0.19,8.09±0.28 and 36.94±0.30,7.59±0.20 and 35.96±0.72,7.50±0.14 and 34.81±0.27,respectively.There was significant statistical difference between the values of the two groups(t=-84.00,-182.72,-261.27,-186.48,-100.22,-239.00,respectively,P＜0.01).At each time point,inflammatorily hyperplastic nodes had significantly higher values of SNRs compared to tumor-bearing nodes(t=83.97,174.07,158.49,152.71,96.06,154.57,respectively,P＜0.01).A complete or part signal filling defect occurred in the tumor-bearing lymph node on the MR lymphographic images.Conclusions The internal anatomy and function of the lymph nodes were effectively visualized by interstitial MR lymphography with positive lymphotropic contrast agent,which provide a sensitively diagnostic way for the differentiation of benign lymph nodes from malignant ones.