Objective To evaluate the effect of dexmedetomidine alternative to propofol administered in general anesthesia on maintainance and recovery in patients undergoing gynecologic laparoscopy.Methods One hundred and twenty patients undergoing gynecologic laparoscopy,were divided into group C,D1 and D2 with 40 cases each by random digits table method.Group C received propofol 100 μ g / (kg min) and remifentanil 0.10-0.25 μ g / (kg min) intravenous infusion,group D1 received propofol 100 μ g/(kg min),remifentanil 0.10-0.25 μ g/(kg min) and dexmedetomidine 0.2 μ g/(kg h) intravenous infusion,while group D2 received dexmedetomidine 2.0-3.0 μ g / (kg h) and remifentanil 0.10-0.25 μ g/ (kg min).Cis-atracurium was given on time to maintainance of anesthesia.The heart rate (HR),mean arterial pressure (MAP) were recorded at the following time points:arriving at operating room (To),anesthesia induction (T1),intubation (T2),skin incision (T3),pneumoperitoneum (T4),10 min after pneumoperitoneum (T5),20 min after pneumoperitoneum (T6),the end of operation (T7),discharge from post anesthesia care unit(T8) and the first postoperative day (T9).The time of emergence,extubation and duration in post anesthesia care unit were recorded too.Ramsay scale and Riker Sedation-Agitation Scale on emergence,extubation and discharge from post anesthesia care unit and post anesthesia recovery score at T9 were also recorded.Results The MAP in group D2 was higher at T3 to T7 time-points than that in group C and group D1 [(93 ± 10),(99 ± 11),(94 ± 13),(95 ± 10),(91 ± 10) mm Hg (1 mm Hg =0.133 kPa) vs.(88± 11),(90± 10),(87±9),(86±9),(83±8)mmHgand (87±9),(90± 10),(86±8),(85 ±7),(83 ± 7) mm Hg],there were significant difference (P ＜ 0.05).There were no significant difference among groups on the HR at each time point and the time of emergence,extubation and duration in post anesthesia care unit (P ＞ 0.05).Ramsay scale was lower on emergence and Riker Sedation-Agitation Scale was higher on emergence and extubation in group C than that in group D1 and group D2 [(3.7 ± 1.3) scores vs.(4.0 ± 0.9),(4.2 ±0.9) scores,(3.1 ± 1.0) scores vs.(2.8 ±0.6),(2.7 ±0.9) scores,(3.3 ±0.7) scores vs.(3.2 ± 0.4),(3.0 ± 0.5) scores],there were significant differences (P ＜ 0.05).Riker Sedation-Agitation Scale was higher on extubation in group D1 than that in group D2(P ＜ 0.05).Post anesthesia recovery score at T9 in group D2 was apparently increased compared with that in group C and group D1 [(108 ± 10) scores vs.(93 ± 13),(93 ± 15) scores] (P ＜ 0.05).Conclusions Dexmedetomidine 2.0-3.0 μ g/ (kg h) administered in general anesthesia on maintainance in gynecologic laparoscopy can improve the quality of extubation and promote postoperative recovery without prolonging extubation time,but have a influence on hemodynamic changes at early stage of pneumoperitoneum.

Objective To investigate the changes of plasma IL-6 and IL-8 level and the inhibitory effects of preoperative administraction of bile salt or anti-endotoxin antibody against endotoxin in cases of obstructuve jaundice.Methods 50 cases with obstructive jaundice were randomized into OJT1 group taking bile salt orally,OJT2 group receiving intravenous anti-endotoxin antibody and OJ group receiving general treatment.The plasma levels of endotoxin(ET),interleukin-6(IL-6) and interleukin-8(IL-8) were determined in all the cases.Results The plasma ET level was significantly lower in the patients of OJT1 and OJT2 groups than in those of OJ group preoperatively and was further decreased postoperatively(P

Cancer/testis(CT) antigens,of which more than 40 kinds have now been identified,are encoded by genes that are expressed normally in the human germ line,and are also expressed in various tumor types,including melanoma,and carcinomas of the bladder,lung and liver.These immunogenic proteins are being vigorously pursued as targets for therapeutic cancer vaccines.In this paper,the classification,expression and function of CT antigens are reviewed.

Objective To observe the changes of mechanical pain thresholds and autophagy related proteins microtubule-associated protein 1 light chain 3 (LC3) and sequestosome 1 (SQSTM1 also known as p62) expression levels in the C57BL/6 mouse models of chronic prostatitis/ chronic pelvic pain syndrome (CP/CPPS),and provide animal experimental evidence for CP/CPPS pain and autophagy study.Methods 36 male C57BL/6 mice were randomly divided into three groups: the model group,control group and na(i)ve group.The CP/CPPS model was established by subcutaneous injection in the lower abdomen region with suspension liquid,containing protein extract of male SD rat prostate gland and complete Freund adjuvant.At 1month and 6 months after modeling,the mice were sacrificed and prostate tissues were harvested for histological examination using HE staining.Mechanical tactile hyperalgesia was measured with von Frey filaments.The autophagy-related proteins LC3 and p62 expression levels were detected by immunohistochemistry,respectively.The average IOD was measured by Image Pro Plus 6.0,and the statistical analysis was performed with GraphPad Prism 5 software.Results The histopathology showed the appearance of chronic prostatitis in the model group,representing hyperplasia and lymphocytic infiltration to a different degree and lasted for 6 months after modeling.Moreover,prostate intraepithelial neoplasia (PIN) appeared in the model group at 6 months after modeling,characterized by the disappearence of basement membrane and obvious nuclear abnormality,while the control and na(i)ve groups showed normal histology during the 1-6 months.Compared with the control and na(i)ve groups,the mechanical pain threshold in the model group was significantly decreased along with the time from (0.353±0.154) g at 0 week to (0.008±0.00) g at 22 weeks (P<0.05).The average IOD of LC3 and p62 expression in the model group was significantly increased with timing from [(2.767±0.464)%,(2.872±1.642)%] at 1month to [(13.501±1.900)%,(9.07±0.49)%] at 6 month,P<0.05.Conclusions A CP/CPPS model is successfully established in C57BL/6 mice.For the model group,the mechanical pain threshold is decreased and autophagy levels are increased gradually with time.These phenomena show that chronic inflammation microenvironment may promote pain and autophagy activity in the prostate,which is closely related with the occurrence and development of prostatic intraepithelial neoplasia.

ObjectiveTo evaluate whether AN69 ST membrane would prolong filter lifetime in continuous renal replacement therapy (CRRT) without anticoagulation in patients with high risk of bleeding.Methods A single-center, prospective, randomized, double-blind control trial with crossover design was conducted. From March 1st to December 31st in 2013, patients who were admitted to Department of Critical Care Medicine of the Fourth Hospital of Hebei Medical University meeting CRRT treatment indications, but could not receive systemic anticoagulation because of high risk of bleeding were studied. The selected patients were randomly divided into two groups according to a random number table, and four filters consisting of two AN69 ST100 membrane filters (A) and two traditional AN69 M100 membrane filters (B) were used for them. GroupⅠ with the filter order of A-B-A-B, and groupⅡ with the order of B-A-B-A. The clinical data of patients was recorded in detail, and conventional AN69 ST and AN69 membrane filter lifetime, their influence on coagulability, and the incidence of bleeding complications were compared.Results Seventeen patients were enrolled, with 10 in groupⅠ, and 7 in groupⅡ. The basic medical characteristics including gender, age, acute physiology and chronic health evaluationⅡ (APAECHⅡ) score, sequential organ failure score (SOFA), Acute Renal Injury Network (AKIN) stage, activated partial thromboplastin time (APTT), prothrombin time (PT), international normalized ratio (INR), platelet count (PLT), and use of mechanical ventilation were not significantly different between two groups. But the use of vasoactive drug was more frequent in groupⅡcompared with that of groupⅠ[100.0% (7/7) vs. 30.0% (3/10),χ2 = 8.330,P = 0.010]. AN69 ST filter lifetime (n =34) was (15.92±2.10) hours, there was no statistically significant difference compared with that of AN69 membrane (t = 0.088,P = 0.942), filter lifetime of which (n = 34) was (16.12±1.38) hours. It was also found by Kaplan-Meier survival analysis that there was no significant difference between the two membrane filter lifetime (χ2=1.589,P =0.208). Logistic regression analysis showed that the life of the first filter was not correlated with coagulation indicators, including APTT, PT, INR, and PLT [APTT: odds ratio (OR) = 0.977, 95% confidence interval (95%CI) = 0.892-1.071, P = 0.623; PT:OR = 1.001, 95%CI = 0.901-1.109,P = 0.988; INR:OR = 1.078, 95%CI = 0.348-3.340,P = 0.896;PLT:OR = 0.996, 95%CI = 0.974-1.019,P = 0.735]. The application rate of vasoactive drugs, which was different between two groups for basic medical indications showed no effect on filter life time (OR = 2.541, 95%CI = 0.239-26.955,P = 0.439). Reasons of clotting in filters were also analyzed, and it was found that blood coagulation in the filter ranked the top (88.2%), and the other reasons were catheter-related problems, death, and unscheduled transport. No difference in blood coagulation function was found in both groups after treatment for 12 hours, and there was no bleeding complication.ConclusionDuring the CRRT without systemic anticoagulant, both surface-treatment with polyethyleneimine AN69 and AN69 ST membrane cannot prolong filter lifetime.

Objective To investigate the feasibility of using clustered regularly interspaced short palindromic repeats ( CRISPR)-mediated genome editing to downregulate the expression of programmed cell death protein 1 (PD-1) on primary T cells by using a lentivirus delivery system. Methods Lentivirus vec-tors pLentiCRISPR A1-A6 containing different PD-1 genomic DNA sequences as single guide RNA ( sgRNA) for Cas9 targeting were constructed individually. The lentivirus vectors were tranduced into primary CD4 T cells. Flow cytometry analysis was performed to detect the expression of PD-1 for evaluating the knockout ef-ficiency. Results The lentivirus vectors pLentiCRISPR A1-A6 carrying six different target sites were con-structed and respectively tranduced into primary CD4 T cells. The expression of PD-1 accompanied with the activation of T cells. Co-expression of CD25 and PD-1 was observed on activated T cells. All of the six sites could be targeted by Cas9, of which A2 and A6 sites were more efficient in knocking out the gene encoding PD-1 with a rate of 19% and 29%, respectively. Conclusion This study suggests that it is feasible to knock out the expression of PD-1 on primary T cells by using CRISPR.

BACKGROUND: Telomerase can maintain the telomere length and avoid cel replicative senescence and apoptosis in somatic cells. Its catalytic subunit cal ed telomerase reverse transcriptase has roles in mediating cellsurvival and anti-apoptotic functions. OBJECTIVE: To evaluate the effects of human telomerase reverse transcriptase on amyloid β1-40-induced human embryonic cortical neurons injury. METHODS: Human cortical neurons derived from 12-16 weeks old aborted fetuses were transfected with recombinant adenovirus vector encoding human telomerase reverse transcriptase. Expression of human telomerase reverse transcriptase was evaluated by immunocytochemical staining. Telomerase activity was measured using a PCR-based telomeric repeat amplification protocol enzyme-linked immunosorbent assay kit. Human embryonic cortical neurons were treated with 10 μmol/L ol/L amyloid β1-40 after transfected for 3 days. Cel viability, reactive oxygen species levels and glutathione contents in human embryonic cortical neurons were respectively detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate and chromatometry. RESULTS AND CONCLUSION: Expression of human telomerase reverse transcriptase reached peak at 3 days after transfection, and the telomerase activity was rebuilt; 10 μmol/L amyloid β1-40 could significantly reduce the cel viability of neurons and glutathione content (P < 0.05 and P < 0.01), and increase the reactive oxygen species levels (P < 0.05). The neurons transfected with human telomerase reverse transcriptase gene could be significantly against the toxicity of amyloid β1-40 and increase the cel viability and glutathione content (P < 0.05 and P < 0.01), and decrease the reactive oxygen species levels (P < 0.05). The results indicate that human telomerase reverse transcriptase can protect amyloid β1-40-induced human embryonic cortical neurons injury

Objective:To study whether autoimmune regulator (Aire) affects macrophage polarization. Methods: The mouse mononuclear macrophage cell line RAW264. 7 cells, stable expressing GFP-Aire protein RAW264. 7 cells ( A33-3 ) and stable expressing GFP protein RAW264. 7 cells (C1-6) were stimulated with LPS,IL-4 and LPS combined with immune complex respectively to make the macrophage polarize to M1 (LPS),M2a (IL-4) and M2b (LPS with immune complex). To investigate the effects of Aire on the various types of macrophages polarization,the M1 related molecules (IL-1α,iNOS and IL-6),M2a related molecules (Arg-1) and M2b related molecule (IL-10) were detected by Real-time PCR. Results:The expression of IL-1α,IL-6 and inducible nitric oxide synthase ( iNOS) ,the products of M1 macrophages, were significantly upregulated in RAW264. 7 cells treated with LPS at 0. 5 μg/ml. The expression of Arg-1 and IL-10 mRNA in RAW264. 7 cells increased in a dose-dependent manner after stimulation with IL-4 or LPS combined with immune complexes, respectively. M1 macrophage-related marker iNOS and IL-1 increased, whereas IL-6 levels decreased in A33-3 cells compared with C1-6 cells after treatment with LPS. The expression of Arg1 and IL-10 were downregulated in A33-3 cells compared with C1-6 cells after IL-4 and LPS combined with immune complexes stimulation,respectively. Conclusion:Aire may promote M1 macrophage polarization while inhibit M2a and M2b macrophage polarization.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

Objective To investigate the relation between positive fluid balance and the prognosis in severe sepsis patients with acute kidney injury (AKI).Methods A retrospective analysis of clinical data of 90 patients in our department was carried out.According to the final outcome,patients were divided into survival group (n =26) and non-survival group (n =64),in which the relation between patients' positive fluid balance and the prognosis was evaluated.Results (1) There was no statistically significant difference in the age and severity between survival group and non-survival group of patients with severe sepsis and consequent AKI,but the patients in non-survival group had greater volume overload.Compared with the survival group,higher mean fluid balance [(1 112.12±546.85) mLvs.(644.69±474.93) mL,P=0.00],and less urine output [(1 224.07 ± 708.79) mL vs.(2 032.36 ± 723.53) mL,P =0.00) in non-survival group.(2) There was no significant difference in mortality between early and late continuous renal replacement therapy (CRRT) during ICU care.However,the average daily fluid load in late CRRT patients was significantly greater than that in early CRRT patients [(1178.81 ±397.03) mLvs.(287.22 ± ± 433.53) mL,P =0.00] and the lung oxygenation index in late CRRT patients was significantly worse thanthat in early CRRT patients [(211.22±42.56) vs.(169.46±57.40),P=0.04] (3) The relevant variables to 28-day mortality in AKI patients with severe sepsis included CRRT treatment,oxygenation index and the average daily fluid balance ＞ 500 mL.Among them,fluid balance ＞ 500mL was an independent risk factor for AKI patients with severe sepsis.Their prognosis was worse if they had greater positive fluid balance.CRRT was the protective factor which could affect the prognosis of patients with severe sepsis complicated by AKI.Conclusions Patients with severe sepsis complicated by AKI has a high mortality.Persistent fluid overload can lead to increased mortality in patients with severe sepsis complicated by AKI.Early CRRT can reduce fluid retention in patients with renal failure and improve oxygenation index.