Objective To prepare urokinase plasminogen activator (uPA)-loaded anionic lipid microbubbles (uPA-MBs) for thrombolysis with low-frequency ultrasound in vitro.Methods Anionic microbubbles composing of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC),1,2-dipalmitoyl-sn-glycero-3-phospho-(l'-rac-glycerol) (DPPG),1,2-distearoyl-sn-glycero-3-phosphoethanol amine-N (succinyl PEG2000) (DSPE-PEG2000) and perfluoropropane (C3F8) were prepared by the mechanical vibration method.Then,the resulting anionic microbubbles were incubated with uPA.uPA-MBs were obtained via electrostatic adsorption.Bubble size and distribution were measured by particle size analyzer.FITC-labeled uPA-MBs were obtained and observed under fluorescence microscope.The surface potential of uPA-MBs and plain microbubbles (P-MBs) were detected by Zeta potential analyzer.Sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) was used for confirming the binding of uPA protein and anionic microbubbles.The encapsulation efficiency of uPA-MBs was determined by bicinchoninic acid (BCA) protein assay kit under three different dosages of uPA (10 000,50 000 and 100 000 U).The thrombolysis efficiency of uPA-MBs combined with low-frequency ultrasound was examined in vitro.Two-sample t test,one-way analysis of variance and Bonferroni test were performed to analyze the data.Results UPA-MBs were successfully obtained with the mean particle size of (1.76±0.29) μm.The surface potential of these bubbles was significantly higher than that of P-MBs:(-36.64±0.21) mV vs (-66.33±2.38) mV (t =21.538,P＜0.05).Fluorescence microscope showed a green shell of FITC-labeled uPA-MBs.The encapsulation efficiency of uPAMBs with the added dosage of 10 000 U was (42.01±2.02) ％,which was significantly higher than those of 50000 and 100 000 U ((33.24±1.95)％ and (33.10±1.65)％ respectively,F=22.340,P＜0.05).The thrombolysis efficiency by saline was (4.09±0.80)％,saline + ultrasound (8.50±1.48)％,MBs + ultrasound (14.27± 1.59) ％,uPA-MBs + ultrasound (35.72±6.31) ％ and uPA (16.87±0.46) ％,respectively (F =48.783,t =-8.613,-7.273,-5.942,-6.908,all P＜0.05).Conclusion Anionic microbubbles can successfully load uPA,and achieve significantly better thrombolysis effect when combined with low-frequency ultrasound.

Abstract: Objective　To analyze the etiological characteristics and drug resistance of patients with bloodstream infection (BSI) in the bacterial resistance monitoring network in Hainan Province from 2018 to 2020, so as to provide laboratory data for clinical diagnosis and treatment. Methods　The clinical data of the subjects were collected, and the etiological characteristics of BSI patients and drug resistance of commonly used drugs in clinical treatment were analyzed retrospectively. SPSS 26.0 software was used for statistical analysis. Results　A total of 877 strains were isolated, including Gram-negative bacteria (584 strains, 66.6%), Gram-positive bacteria (239 strains, 27.2%) and fungi (54 strains, 6.2%); male patients (591 cases, 67.4%), female patients (286 cases, 32.6%); inpatients (780 cases, 88.9%), outpatient and emergency patients (97 cases, 11.1%); the main primary diseases of BSI patients were hypertension, cerebral infarction and type 2 diabetes, and the main primary infections were pulmonary infection and urinary system infection. Intensive care unit (25.2%, 221 cases), emergency department (10.9%, 96 cases), oncology department (9.1%, 80 cases), nephrology department (6.8%, 60 cases) and hepatobiliary and pancreatic surgery department (4.3%, 38 cases) had the highest proportion of pathogenic bacteria. Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, coagulase-negative Staphylococcus, Viridans group streptococci and Candida albicans were the most frequently isolated pathogens. The detection rates of carbapenem-resistant Klebsiella pneumoniae, carbapenem-resistant Pseudomonas aeruginosa and carbapenem-resistant Acinetobacter baumannii were 3.4%, 15.2% and 36.4% respectively. The carbapenem-resistant Escherichia coli was not checked out. The detection rates of methicillin resistant Staphylococcus aureus and methicillin resistant coagulase negative Staphylococcus were 18.5% and 79.1% respectively. Conclusions　Gram-negative bacteria are the most common pathogens of BSI, and inpatients are the main source of BSI. Age, underlying diseases and primary infection are the risk factors of BSI. Clinical laboratories should strengthen the etiological monitoring of high-risk patients with BSI, and the resistance analysis of common antibiotics can provide a basis for the rational use of antibiotics in clinical practice.

OBJECTIVE:To evaluate therapeutic efficacy of Xuebijing injection in the treatment of severe craniocerebral trau-ma. METHODS:67 patients with severe craniocerebral trauma selected from neurosurgery department of our hospital were Treat-ment method divided into treatment group(33 cases)and control group(34 cases). Observation group was given conventional treat-ment,i.g. oxygen inhalation,dehydration,nourish cranial nerve,anti-infection. Treatment group On the basis of the control group was given Xuebijing injection 50 ml/time,tid,ivgtt. PT,TT,PLT,FIB,CK,LA, D-D,blood gas index (PaCO2,PaO2, HCO3-)of 2 groups were observed after 7 days of treatment,and prognosis of 2 groups were evaluated after 6 months as well as therapettin efficacy. RESULTS:After treatment,PT,TT,PLT,FIB,CK,LA, D-D,PaCO2,PaO2 and HCO3- of 2 groups were all better than before,and the treatment group was better than the control group,with statistical significance (P<0.05). The rate of good prognosis in treatment group (78.79%) was significantly better than in control group (55.88%) after 6 months of treat-ment,with statistical significance(P<0.05). CONCLUSIONS:Xuebijing injection in the treatment of severe craniocerebral trau-ma can improve coagulation function,blood gas levels and the inflammatory reaction,and is conducive to improve the patient's prognosis.

Objective To evaluate the clinical efficacy and adverse effects of CAG regimen in treatment of primary, refractory and relapsed acute myeloid leukemia (AML) and high risk myelodysplastic syndrome (MDS), and analyse the factors influencing long-term survival. Methods Sixty-one patients with AML ( primary, n = 27; refractory, n = 18; relapsed, n = 16) and 9 patients with MDS were treated with CAG regimen. Examinations on liver and renal function, electrocardiogram and bone marrow cytology were performed before and after treatment, and adverse effects of CAG were observed. Short-term efficacy was evaluated based on clinical manifestation, peripheral blood and bone marrow cytologic examinations. Patients were followed up, overall survival ( OS) and disease free survival ( DFS) were analysed, and long-term efficacy of CAG regimen was evaluated. The factors influencing long-term survival were analysed by Log-rank test of survival curve. Results After a course of treatment by CAG regimen, the total effective rate was 71% , and 34 patients (49%) experienced complete remission. The median time of follow up was 45 months, the median OS was 28 months, and the median DFS was 23 months. Age, level of lactate dehydrogenase (LDH), remission condition after a course of treatment by CAG regimen and adoption of HD-Ara-C regimen as consolidation treatment were influencing factors for OS and DFS. The dominant clinical adverse effects were bone marrow depression, with 13 d as the median duration of agranulocytosis ( neutrophil <0.5 ×10~9/L) and 9 d as the median duration of thrombocytopenia (platelet <20 ×10~9/L). Conclusion CAG regimen may lead to favourable therapeutic effects in treatment of primary, refractory and relapsed AML and high risk MDS, and may yield less adverse effects and better long-term therapeutic effects. Age, level of LDH, remission condition after a course of treatment and adoption of HD-Ara-C regimen as consolidation treatment are dominant influencing factors for survival.

Objective To discuss the histology change,apoptosis state and Bcl-2,Bax expression after the bone cement leakage into the intervertebral disc in vertebroplasty.Methods Eight majority canis familiaris were studied.Three lumbar intervertebral discs(L2 to L5)in each dog were randomly classified into three groups(control group,PMMA group,and CPC group),the canine discs were stabbed by 18-gauge needle,and 0.1 ml cement was injected into them.Control discs were only stabbed and injected with nothing.Histology of all discs was studied 24 weeks after the operation.Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and immunohistochemisty were used to detect apoptosis and Bcl-2,Bax expression in the discs.The data were statistically analyzed by SPSS 12.0.Results Intervertebral disc degeneration was not found in control groups.In bone cement groups,however,ruptured or serpentine patterned fibers,decreased cellularity of the nucleus pulposus and condensed matrix of the nucleus pulposus were found in histologic results.The Bax protein decreased in the order of control group, CPC group,and PMMA group.However,the Bcl-2 protein increased in the order of control group,CPC group,and PMMA group.The histology grade was significantly different among the three groups under ANOVA analysis(P

Objective To detect the expression of BclGL in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) and to investigate its significance.Methods Peripheral blood was obtained from 20 patients with active SLE (A-SLE),18 patients with inactive SLE (Ⅰ-SLE) and 30 healthy controls.Flow cytometry was performed to calculate the number of PBMCs,flow cytometry combined with annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining to determine the early apoptotic rates of PBMCs,fluorescence-based quantitative PCR and Western blot to measure the expression of BclGL mRNA and protein,respectively.The serum level of interferon (IFN)-α was determined by enzyme-linked immunosorbent assay (ELISA).Data were analyzed by using the software SPSS13.0.Mann-Whitney U-test or Kruskal-Wallis test was used for group comparisons.Pearson correlation coefficient test was applied to evaluate the relationship of BclGL expression with cell apoptotic rate and some clinical parameters.Results The number of PBMCs was significantly lower in patients with A-SLE than in those with Ⅰ-SLE and healthy controls ((0.16 ± 0.06) × 109/L vs.(0.27 ± 0.14) × 109/L and (0.34 ± 0.23) × 109/L,both P ＜ 0.01).Increased apoptotic rate of PBMCs was observed in patients with A-SLE compared with those with Ⅰ-SLE and healthy controls ((22.6 ± 1.1)％ vs.(16.4 ±0.9)％ and (10.1 ± 0.4)％,both P ＜ 0.01),and in patients with Ⅰ-SLE compared with the healthy controls (P ＜0.01).The mRNA and protein expressions of BclGL in PBMCs were both significantly higher in patients with ASLE than in those with Ⅰ-SLE and healthy controls (all P ＜ 0.01).A significant increase was observed in serum IFN-α level in the patients with SLE compared with the healthy controls ((32.5 ± 2.2) μg/L vs.(15.5 ± 1.3) μg/L,P ＜ 0.01).The mRNA expression of BclGL in PBMCs from patients with SLE was positively correlated with the apoptotic rate in PBMCs (r =0.486,P ＜ 0.01),SLE disease activity index score (r =0.496,P ＜ 0.01),titers of antinuclear antibodies (r =0.516,P ＜ 0.01) and serum IFN-o level (r =0.535,P ＜ 0.01),but was negatively correlated with complement C3 level (r =-0.515,P ＜ 0.01).Conclusions The increased expression of BclGL in PBMCs may contribute to the abnormal apoptosis in PBMCs,which in turn takes part in the pathogenesis of SLE.

Based on the octahedral modifiable structures and kinetic inertness, platinum (IV) complexes have become antitumor prodrug candidates to mitigate platinum (II) drug resistance and side effects. The nitrobenzoxadiazole derivative (NBDHEX) can inhibit the activity of glutathione S-transferases (GSTs) and be introduced to conjugate with platinum(II) complexes DN603 and DN604 to yield two platinum (IV) complexes DN603/DN604-NBD. In vitro assays demonstrated that DN603/DN604-NBD could significantly inhibit the proliferation of cisplatin-sensitive A549 and resistant A549/cDDP cancer cells. The uptake of DN603/DN604-NBD in A549/cDDP cells was much higher than that of cisplatin, causing a higher rate of cell apoptosis and a greater ratio of Bax/Bcl-2, the activation of caspase-3 and cleavage of DNA repair enzyme (PARP), inducing the mitochondria-dependent cell apoptosis pathway. DN603/DN604-NBD could induce higher reactive oxygen species (ROS) levels, significantly enhance phosphorylation of histone H2AX on Ser-139 (γ-H2AX) fluorescence intensity and cause a greater degree of DNA double-stranded damage. Studies have shown that GSTs kinase GSTP1 was highly expressed in cisplatin-resistant cancer cells. Moreover, DN603/DN604-NBD targeted GSTP1 to suppress its expression level. By using the ROS scavenger NAC and the c-Jun N-terminal kinase (JNK) inhibitor SP600125 in advance, it was found that DN603/DN604-NBD could significantly increase the number of living cells and decrease the phosphorylation levels of JNK and c-Jun. The results indicated that DN603/DN604-NBD could produce higher levels of ROS to activate JNK-mediated signaling pathway, induce apoptosis in tumor cells to overcome cisplatin resistance. All the animal experiments were approved by Animal Ethics Committee of Southeast University (grant No. 20210303025). In vivo assays confirmed that DN603/DN604-NBD could inhibit the growth of A549 xenograft tumors with nearly no toxicity and fewer side effects. Taken together, DN603/DN604-NBD are investigated as two potential novel platinum (IV) prodrug candidates for anticancer therapy.

Objective To evaluate the outcome of posterior screw-rod fixation system in reduction and internal fixation of atlantoaxial dislocation. Methods A retrospective study was done on 27 patients with atlantoaxial instability including 18 male and 9 female (at age range of 13-51 years, mean 31 years) from January 2007 to May 2009. There were 11 patients with chronic odontoid fractures, five with isolated bone odontoid, seven with transverse ligament rupture of atlas and four with rheumatoid arthritis. Skeletal reduction was performed in all the patients. The anterior atlantodens interval (ADI)ranged from 8 mm to 15 mm, average 11 mm. All the patients underwent an intraoperative reduction by posterior C1 lateral mass and C2 pedicle screws with rod fixation. According to American Spine Injury Association (ASIA) impairment scale, there were eight patients at grade B, 15 at grade C and four at grade D. Results All patients were followed up for 6-24 months (average 13 months), which showed that the neck symptoms were improved, with bony union. The ADI was reduced to 2-4 mm (average 2.8 mm)postoperatively. Postoperative ASIA scale was grade C in four patients, grade D in 12 and grade E in 11.There were no neurologic or vascular complications occurred, or no failure of the internal fixtors, pseudarthrosis or instability. Conclusion Posterior screw-rod fixation system has advantages of simple procedures, few complications and good results and can be used for intraoperative reduction of atlantoaxial dislocation.

Objective To investigate the safety of rituximab combination chemotherapy in the treatment of B-cell non-Hodgkin' s lymphoma (B-NHL) complicated with hepatitis B virus (HBV) infection,and assess the incidence of HBV reactivation reduced by prophylactic lamivudine.Methods A retrospective study of HBV-related markers,HBV-DNA and liver function was performed before and after rituximabcontaining treatment in B-NHL patients.Thirty nine B-NHL patients with HBcAb(+)/HBsAb(-) were divided into prophylactic group (14 cases) and control group (25 cases).The incidences of HBV reactivation,functional damage of liver were measured.Results Among the 108 B-NHL patients who received rituximab combinatio nchemotherapy,15 (13.89 ％) were HBsAg (+) and 39 (36.11％) HBsAg (-) / HBcAb (+).Of the 15 HBsAg (+)patients,2 (13.3 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 7.7 ％(1/13) in patients who received prophylactic antiviral treatment and 50 ％ (1/2) in those who did not receivelamivudine.Among the 39 HBsAg (-) / HBcAb (+) patients,3 cases (7.7 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 0 in patients who receivcd prophylactic lamivudine treatment and 12 ％ (3/25) in those who did not receive this antiviral drug.Conclusion Prophylactic lamivudine before rituximab combination chemotherapy can reduce HBV reactivation obviously.

The complete genomic sequence of Duck hepatitis virus 1 (DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides (nt) in length with a 5'-terminal un-translated region (UTR) of 626 nt and a 3'-terminal UTR of 315 nt (not including the poly(A) tail). One large open reading frame (ORF) was found within the genome (nt 627 to 7 373) coding for a polypeptide of 2 249amino acids. Our data also showed that the poly (A) tail of DHV-1 has at least 22 A's. Sequence comparison revealed significant homology (from 91.9% to 95.7%) between the protein sequences of the virus in the Picornaviridae family, its genome showed some unique characteristics. DHV-1 contains 3copies of the 2A gene and only 1 copy of the 3B gene, and its 3'-NCR is longer than those of other picornaviruses. Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates (from 92.9% to 99.2%), indicating that DHV-1 is genetically stable.