Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .

Objective To explore the dynamic changes of von willebrand factor(VWF)and Pseleetin in the finger-replanted patients,and the relationship between the prognosis of the surgery and hypercoagulability.Methods From December 2004 to December 2006,eishty finger-replanted patients were recruited to our study.with 40 healthy volunteers as controls.Plasma VWF and P-selectin were detected by enzyme-linked immunosorbent assay(EUSA)in both controls and patients before or after replantation.Results The VWF and P-selectin levels had significant differences between the replantations and controls(F=14.76,11.76,P＜0.01).The VWF levels in the patients of 1,4,8,16 hours after replantation were(1 715±493),(1 396±549),(1 266±504),(1 163±436)U/L respectively,all markedly higher than the controls(P＜0.01).The P-selectin levels in patients of 1,4,8,16,24 hours after operation were(14.7±2.6),(12.5±3.0),(11.8±3.2),(11.1±3.0)、(10.5±2.6)μg/L,significanfly higher than the controls(P＜0.01).The VWF levels in patients of pre-replantion and the 1,4,8,16,24,48,72 hours after replantation were(854±209),(1 535±389),(1 177±407),(1 040±283),(958±216),(829±193),(777±151),(713±137)U/L in successful group,and were(1 202±164),(2 333±243),(2 146±161),(2 039±244),(1 865±170),(1 645±283),(1 427±331),(1 188±262)U/L in unsuccessful groups.They were all significantly different at the same test-time points between two groups(t=4.44,5.12,6.10,8.43,10.17,8.85,5.10.4.61,P＜0.05).The P-selectin levels in patients of 1,4,8,16,24,48,72 hours after replantation were(13.9±2.5),(11.2±2.0),(10.2±1.6),(9.6±1.2),(9.2±0.9),(9.5±0.6),(9.3±0.4)μg/L in successful group,and(17.2±1.0),(16.9±1.0),(17.0±1.3),(16.1±1.1),(14.9±1.5),(13.8±1.4),(12.8±1.2)μg/L in unsuccessful group.Significant difference existed at the same testtime points between two groups again(t=5.22.9.91,10.35,12.79,9.46.9.45,9.33,P＜0.01).After replantation,both VWF and P-selectin were rapidly elevated and went to the summit 4 hours later,then declined to pre-replantation level about 24 to 48 hours later after replantation.Conclusions VWF and P-selectin were associated with the hypercoagulability.Dynamic monitoring VWF and p-selectin may be useful in determining the existence of hypercoagulability and the therapy of anti-coagulability.

Objective To perform a meta-analysis on the associations of interleukin (IL)-6-174G ＞ C (rs1800795) and-634C ＞ G (rs1800796) polymorphisms with type 2 diabetic nephropathy (DN).Methods The data on the studies about the associations of IL-6-174G ＞ C and-634C ＞ G polymorphisms with type 2 DN were collected from Pubmed,Embace,CNKI,Wan Fang and VIP database during their inception and April 2017.The statistical analysis was performed with STATA 14.0 and Review manager 5.3 softwares.The heterogeneity in the eligible studies was assessed by Q-statistic and I2 statistic.When the significant heterogeneity was found,the random effect model was used for meta-analysis,otherwise,the fixed effect model was used.The publication bias was evaluated with funnel and Begger graphs.The pooled odds ratios (OR) and corresponding 95％ confidence intervals (95％ CI) were calculated for evaluating the associations of IL-6-174G ＞ C and-634C ＞ G polymorphisms with type 2 DN.In addition,the sub-group analysis was performed according to the regions of subjects.Results A total of 11 studies were enrolled,The studies on the association of IL-6-174G ＞ C polymorphism with type 2 DN included 1 688 subjects,while those on the association of IL-6-634C ＞ G with type 2 DN included 2 180 subjects.In the association analysis of IL-6-174G ＞ C polymorphism with type 2 DN of Asian population,the significant relationship was detected in an allelic genetic model (OR =0.461,95％ CI:0.274-0.777,P ＜ 0.01),a homozygote model (OR =0.126,95％ CI:0.022-0.734,P =0.021),a recessive genetic model (OR =0.146,95％ CI:0.026-0.827,P =0.030) and a dominant genetic model (OR =0.504,95 ％ CI:0.273-0.930,P =0.028),but not in a heterozygote model (OR =0.606,95 ％ CI:0.321-1.143,P =0.122).There was no significant relationship between IL-6-174G ＞ C polymorphism and type 2 DN in European population.In the association analysis of IL-6-634C ＞ G polymorphism with type 2 DN of Asian population,the significant relationship was found in an allelic genetic model (OR =1.467,95％ CI:1.238-1.737,P ＜0.01),a homozygote model (OR =2.793,95％ CI:1.844-4.230,P =0.021),a recessive genetic model (OR =2.296,95 ％ CI:1.586-3.323,P ＜ 0.01) and a dominant genetic model (OR =1.377,95％CI:1.109-1.711,P ＜ 0.01),but not in a heterozygote model (OR =1.733,95％ CI:0.932-1.476,P =0.174).There was no significant relationship between IL-6-634C ＞ G polymorphism and type 2 DN in European population.Conclusion In Asian population,IL-6-174CC genotype may prevent the progression of type 2 DN,however,IL-6-634GG genotype may promote the development of type 2 DN.But in European population,there is no relationship between IL-6-174G ＞ C and-634C ＞ G polymorphisms and type 2 DN.

Objective To analyze the blood perfusion characteristics of retroperitoneal benign and malignant tumors by real-time contrast enhanced ultrasonography and discuss its value in the differential diagnosis.Methods The study involved 42 patients with pathological evidence through surgery or needle biopsy,including 12 patients with benign retroperitoneal tumors and 30 patients with malignant retroperitoneal tumors.The blood perfusion characteristics of two groups were observed under low mechanical index after the injection of contrast ultrasound agent(SonoVue),and the changes of twodimensional ultrasonography and the time-intensity curve(TIC)were analyzed. Results During real time contrast enhancement,the different characteristics of retroperitoneal benign and malignant tumors were observed.Malignant tumors in retro peritoneum presented the pattern of ultrasound contrast agent(UCA) enhancement from center to periphery and enhanced overall uneven mainly,begin tumors presented peripheral enhancement pattern or uniformity and overall strengthening of the main.TIC curve between benign and malignant tumors displayed that contrast enhanced intensity of region of interest(P＜0.00 1),ascending slop and halftime of descending were statistically significant(P＜0.05).Conclusions Real-time contrast-enhanced ultrasonography is a valued method to provide information for the differential diagnosis in retroperitoneal benign and malignant tumors.

This paper introduces the concepts related to sexual health and sexual health care, summarizes the contents, application scope and limitations of sexual health care assessment tools for oncology nurses at home and abroad, analyzes the problems existing in the assessment tools and puts forward suggestions, aiming at providing theoretical reference for the localization development of sexual health care assessment tools and the development of sexual health care.

Objective:To search for circulating complement-related proteins that predict hypertensive disorders of pregnancy (HDP) based on reports of the development of gestational hypertension and proteinuria and to investigate the role of the complement system in the development of HDP.Methods:A nested case-control study was used, the serum samples of pregnant women who had been given birth or cesarean section in Guangzhou Women and Children′s Medical Center from November 2014 to March 2017 were collected. A total of 60 HDP and 60 normal pregnant women were included and matched 1∶1 by age and gestational week. Unlabeled mass spectrometry was used to screen the differential expression of complement factors in serum samples of 12 pairs of HDP patients and normal pregnancy collected before 20 weeks of pregnancy, and another 48 pairs of serum samples of HDP patients and normal pregnant women were used for preliminary verification. It was selected when the fold change (FC) of complement factor expression was>1.2 or <0.8 and P<0.05. ROC curve was used to evaluate the diagnostic value of corresponding factors. Results:FC of serum C1s, C8 beta chain (C8β) and C1 inhibitor (C1-INH) of HDP patients were 1.19, 1.23, 0.73 ( t=2.07, 2.06, -3.40; P<0.05), respectively. FC of serum C1s, C8 β, C1-INH, factor H-related protein 5 (CFHR5), clusterin (CLU), and C-reactive protein (CRP) of PE patients were 1.39, 1.50, 0.72, 2.49,4.38, and 1.82 respectively ( t=4.36, 5.61, -3.70, 6.82, 8.70, 7.27; P<0.05).The AUC of combining C1s, C8 β and C1-INH was 0.89 in HDP. The AUC of CFHR5, CLU, and CRP in preeclampsia was 0.88, 0.92, and 0.91. Conclusions:Before HDP, the activation and regulation of classic complement pathway and alternative pathway were disordered in pregnant women. The combined detection of complement C1s, C8 β and C1-INH is expected to be used in the prediction of HDP, and CFHR5, CLU, and CRP are expected to be used in the prediction of preeclampsia.

Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.

Objective To investigate the feasibility and safety of using catheter fenestration technology in transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC)supplied by the right inferior phrenic artery(RIPA).Methods From March 2023 to May 2023,five HCC patients,whose HCC lesions were supplied by RIP A,received TACE by using catheter fenestration technology to accomplish the superselective catheterization of RIPA after the conventional microcatheter catheterization of RIPA failed.Results Superselective catheterization of RIPA and TACE were successfully accomplished in all the 5 patients,with a surgical success rate of 100％.The time spent for superselective catheterization of RIPA was(3.2±2.39)minutes.After TACE,the levels of the tumor indicators were decreased when compared with the pre-TACE values,while the liver function indexes showed no obvious damage.Based on the mRECIST criteria,PR was obtained in 3 patients and CR was obtained in 2 patients.None of the five patients developed serious adverse reactions.Conclusion In treating patients with HCC supplied by RIPA,the use of catheter fenestration technology can safely and effectively improve the success rate of superselective catheterization of RIPA during TACE.

Inflammation plays an important role in the development of acute lung injury (ALI). Severe pulmonary inflammation can cause acute respiratory distress syndrome (ARDS) or even death. Expression of proinflammatory interleukin-1β(IL-1β) and inducible nitric oxide synthase (iNOS) in the process of pulmonary inflammation will further exacerbate the severity of ALI. The purpose of this study was to explore the effect of Palrnatine (Pa) on lipopolysaccharide (LPS)-induced mouse ALI and its underlying mechanism. Pa, a natural product, has a wide range of pharmacological activities with the potential to protect against lung injury. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to detect the expression and translation of inflammatory genes and proteins in vitro and in vivo. Immunoprecipitation was used to detect the degree of P65 translocation into the nucleus. We also used molecular modeling to further clarify the mechanism of action. The results showed that Pa pretreatment could significantly inhibit the expression and secretion of the inflammatory cytokine IL-1β, and significantly reduce the protein level of the proinflammatory protease iNOS, in both in vivo and in vitro models induced by LPS. Further mechanism studies showed that Pa could significantly inhibit the activation of the protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway in the LPS-induced ALI mode and in LPS-induced RAW264.7 cells. Through molecular dynamics simulation, we observed that Pa was bound to the catalytic pocket of Akt and effectively inhibited the biological activity of Akt. These results indicated that Pa significantly relieves LPS-induced ALI by activating the Akt/NF-κB signaling pathway.