A benzyl functionalized ionic liquid, 1-benzyl-3-methylimidazolium bis [( trifluoromethyl ) sulfonyl]imide ([BeMIM][Tf2 N]), was synthesized and characterized as an extraction solvent of dispersive liquid-liquid microextraction ( DLLME) for enrichment and determination of 5 organophosphorus pesticides (phoxim, fenitrothion, chlorpyrifos, phorate and parathion) and 2 aromatic compounds (chloronaphthalene and anthracene) from environmental water samples by high-performance liquid chromatography ( HPLC). [BeMIM] [ Tf2 N] had higher extraction efficiency than 1-octyl-3-methylimidazolium bis [( trifluoromethyl) sulfonyl]imide and common organic solvents such as CCl4 and C2 Cl4 . The extraction was performed using 40 μL of [BeMIM][Tf2N] and 1 mL of methanol as extraction solvent and dispersive solvent respectively with centrifugal time of 5 min. Under the optimal conditions, the method proposed here provided a good linearity for all analytes with correlation coefficients between 0. 9994 and 0. 9998. The repeatability values, described as intra-day and inter-day relative standard deviations (RSDs) of five replicate experiments at three different concentrations of 10, 40 and 100 μg / L, were 1. 1% -4. 3% and 0. 8% -4. 8% , respectively. The limits of detection (LOD) were 0. 01 μg / L-1. 0 μg / L at a signal-to-noise ratio (S / N) of 3. This developed method was convenient and speedy, and could be employed to detect the analytes in three real environmental water samples with satisfactory relative recovery of 82. 7% -118. 3% and RSD of 0. 7% -5. 6% . Introduction of benzyl group into the imidazolium could obviously enhance the extraction efficiecny for analytes due to the π-πinteraction between [BeMIM] [ Tf2 N] and analytes. [ BeMIM] [ Tf2 N] was a satisfactory extraction solvent with a high enrichment factor of 339 and extraction efficiency of 81. 4% . Partition coefficients of all analytes in [BeMIM][Tf2 N]-DLLME system were determined and the extraction mechanism was discussed.

Objective To explore the reproductive outcomes of different management strategies of ectopic pregnancy .Methods A retrospective cohort study was performed for 648 consecutive patients with a diagnosis of ectopic pregnaney between June 2011 and september 2013.The follow-ups were conducted telephone interviews of pregnancy outcomes after various treatments .Results The subsequent intrauterine pregnancy rates after surgical , medical and expectant managements were 89(65.0%), 35(79.6%) and 6 (54.5%) the intrauterine pregnancy was higher for medical treatment than that for surgical treatment (P<0.05).The subsequent infertility rate after surgical , medical and expectant management were 4.5%, 36.4%, 19.0%.The subsequent infertility rate after medical treatment was lower than that for surgical and expectant managament ( P<0.05) .The re-ectopic pregnancy rates of laproscopic versus abdominal operation were [11 (11.7%), 13(30.2%)] with significant difference (P<0.05).And abdominal operation was higher than laproscopic operation .Conclusion For younger ectopic pregnancy patients , medical treatment shold be as possible as we can .If operation is necessary , laproscopic operation is preferred .

Objective To systematically review the effect of ω-polyunsaturated fatty acids-supplemented nutrition support on the immune function and nutritional status of patients with gastrointestinal malignancies.Methods Databases of MEDLINE,EMBASE,Cochrance Central register of controlled trials,China National Knowledge Infrastructure (CNKI),China Biology Medicine disc (CBM) were searched.The search words were omega-3 polyunsaturated fatty acids,fish oil,cancer,turmor,neoplasms in English or Chinese.All the high quality randomized controlled trials were included after assessed with Jadad scale.RevMan 5.2 was used for statistical analysis.Results Altogether 12 studies were included.Meta-analysis results showed that CD4+％ (I2 =0％,95％ CI=3.44～6.93,Z=5.82,P＜0.00001),CD8+％ (I2 =0％,95％ CI=2.44～7.13,Z=4.00,P＜0.000 1),CD4+％/CD8+％ (I2 =74％,95％ CI=0.16～0.83,Z=2.87,P=0.004),IgA (I2=14％,95％ CI=0.21 ～0.43,Z=5.84,P＜0.00001),prealbumin (I2 =0％,95％CI =0.02 ～ 0.05,Z =3.94,P ＜ 0.000 1) in the treatment group were higher than in the control group,C-reactive protein (I2 =76％,95％ CI =-21.33 ～-5.03,Z =3.17,P =0.002) in the treatment group was lower than in the control group,the differences were all statistically significant.But there were no statistically significant differences in CD3 + ％ (I2 =0％,95％ CI =-3.50 ～ 2.56,Z =0.31,P =0.76),IgG (I2 =92％,95％ CI=-0.32 ～5.54,Z=1.74,P=0.08),IgM (I2 =99％,95％ CI=-0.52～1.89,Z=1.11,P=0.27),total protein (I2 =0％,95％CI=-4.08 ～1.77,Z=0.78,P=0.44),albumin (I2 =29％,95％ CI=-0.87 ～1.45,Z=0.49,P=0.63),transferrin (I2 =0％,95％ CI=-0.23～0.19,Z =0.19,P =0.85) between treatment and control groups.Conclusion Omega-3 polyunsaturated fatty acids can improve the perioperative immune function of patients with gastrointestinal malignant tumor,reduce inflammatory and stress reactions,but have no obvious effect on the improvement of the nutritional status.

Objective To preliminarily understand the infection of Chikungunya virus (CHIKV) in Cangyuan County, a southern border area of Yunnan Province, and provide a reference basis for the prevention and control of Chikungunya fever. Methods In April 2020, a total of 400 serum samples from individuals seeking medical care at the People's Hospital of Cangyuan County in Yunnan Province were collected. Among these, 121 samples were from healthy individuals undergoing physical examinations, and 279 samples were from patients with fever. The serum samples collected underwent CHIKV neutralizing antibody testing using a serum micro-neutralization assay. Real-time fluorescence quantitative RT-PCR was used to detect CHIKV nucleic acid in the samples, followed by analysis of the test results. Results The results of neutralizing antibodies showed that 18 of the 400 human serum samples were positive for neutralizing antibodies against CHIKV, with an overall positivity rate for serum samples of 4.5% (18/400). Among the 279 serum samples collected from patients with fever, 18 were positive for neutralizing antibodies against CHIKV, with a positive rate of 6.45% (18/279), and the neutralizing antibody titers ranged from 1∶10 to 1∶320. The results of 121 healthy human serum samples were negative for neutralizing antibodies against CHIKV. The results of real-time fluorescence quantitative RT-PCR showed that 3 of the 400 human serum samples were positive for CHIKV nucleic acid, and the positive rate was 0.75% (3/400). Among the 279 serum samples collected from patients with fever, 3 samples were positive for CHIKV nucleic acid, with a positive rate of 1.08% (3/279), and Ct values ranged from 36.58 to 37.74. While all healthy human serum samples were negative for CHIKV nucleic acid. Conclusions The findings indicate that infection of CHIKV exists in the population of Cangyuan County, a southern border area of Yunnan Province, and an outbreak of the disease is occurring. Therefore, it is necessary to strengthen the monitoring, prevention, and control of CHIKV in this area.

Objective This study aims to comprehensively investigate the molecular characteristics of the predominant circulating Dengue virus type 1 (DENV-1) during the 2019 Dengue fever outbreak in Xishuangbanna, providing an essential insight to support the prevention and control of dengue fever in the local area. Methods A Dengue virus type 1 (DENV-1) strain, designated as JHS45, isolated from the blood of a febrile patient in Xishuangbanna in 2019, underwent a process of inoculation and cultivation in C6/36 cells. Second-generation sequencing was employed to capture the viral genetic sequence. Bioinformatics software, including CLC, was used for assembling the sequencing data. Sequentially, sequence alignment, construction of a phylogenetic tree, and analysis of amino acid sites were conducted using software such as Lasergene and MEGA6.1. Results Cytopathic effects of JHS45 appeared in C6/36 cells after 6 days. After sequencing and assembly, a 10 687-nucleotide (nt) long sequence of the JHS45 virus was obtained (GenBank accession number: OR593353). Genetic evolutionary analysis revealed that the JHS45 virus formed an evolutionary branch with DENV-1 genotype I viruses prevalent in Xishuangbanna, Guangzhou, Henan, and Zhejiang in China during 2019, as well as the DENV-1 genotype I virus prevalent in Thailand in 2013, with nucleotide homology of 97.6% to 99.9% and amino acid homology of 99.1% to 100%. Further analysis revealed that the JHS45 strain shared a smaller evolutionary branch with the DENV-1 genotype I viruses prevalent in Xishuangbanna (MW386863) and Guangzhou (MW261839) in 2019, showing the highest homology with nucleotide and amino acid homology of 99.9% and 100%, respectively. Amino acid differential site analysis between the JHS45 strain and the DENV-1 prevalent in Xishuangbanna since 2015 revealed 40 amino acid differential sites in the coding region of the JHS45 virus, primarily concentrated in the NS3 and NS5 regions of non-structural proteins. Conclusion The comprehensive analysis of the JHS45 strain's whole genome sequence indicates it is a DENV-1 genotype I virus. The genetic evolutionary relationship between this Xishuangbanna dengue fever outbreak is closely related to the prevalent virus strains in Xishuangbanna, Guangzhou, Henan, and Zhejiang. These findings provide a robust scientific foundation for monitoring dengue fever outbreaks, conducting virus evolution studies, and shaping effective prevention and control strategies, not only within Yunnan Province but also on a broader scale throughout China.

Objective To investigate the clinical features, prognosis and risk factors of patients of acquired immunodeficiency syndrome (AIDS) complicated with cryptococcal meningitis (CM). Methods Totally 26 patients of AIDS with CM who were hospitalized in the No. 8th People's Hospital of Guangzhou were enrolled in this study. The clinical data including diagnosis,experimental and etiological test,treatments and prognosis from all the patients were analyzed retrospectively. The results of cerebrospinal fluid routine test and CD4+ T lymphocyte were compared with those of AIDS patients complicated with tuberculous meningitis. Results Among the 26 patients enrolled in the study, the positive rate of cerebrospinal fluid india ink smear or Crypotococcus neoforrnans euhure was 84.6%. The most common symptoms were fever, headache and meningeal irritation sign. The average CD4+ lymphocyte count was 17.83 × 106/L, which was statistically different from that of tuberculous meningitis patients. All the patients showed concomitant multiple organ infections. The mortality rate was as high as 42.3%. At the end of therapy, the cell counts in the eerebrospinal fluid were remarkably higher in the patients with unfavorable prognosis compared to the patients with good prognosis, which was statistically different. Conclusions CD4+ lymphocyte count is an important marker for differentiating CM from tuberculous meningitis in AIDS patients. The results of cerebrospinal fluid routine test can predict the prognosis.

Objective The purpose of this study was to investigate the value of serum galactomannan (GM) in the diagnosis of invasive pulmonary aspergillosis (IPA) in patients with chronic obstructive pulmonary disease (COPD).Methods We enrolled 60 COPD patients in the study, including 19 IPA and 41 non-IPA cases.We examined serum GM of the patients by ELISA, evaluate the value of serum GM test for the diagnosis of IPA in patients with COPD, and compared the GM values before and after treatment.Results With 0.5 as the positive cutoff value, the sensitivity, specificity, positive predictive value, and negative predictive value of serum GM were 57.9%, 95.3%, 84.6%, and 83.0%, respectively, with a high specificity and a low sensitivity.The 7 IPA cases showed a significantly decreased GM value after treatment as compared with the baseline (0.30±0.21 vs 1.48±1.37, P=0.004).Conclusion The serum GM test has a limited value in the diagnosis of IPA in patients with COPD, but dynamic monitoring of the changes of the serum GM value may help evaluate the patient's condition.

The most difficult field in gene therapy is that vector system should offer both a means of successful transfection and a maximum of safety for the patient. Viral vectors and plasmid vectors are traditional vectors; they may cause unwanted immunological side effects resulting from the expression of nontherapeutic genes. Our aim is to develop a new general gene therapy vector which is suggested to be called as Micro-Linear Vector. The gene expression cassette is capped by our designed cap, including promoter, enhancer, objective gene, and RNA-stabilizing sequence, so it can defend the exnuclease in the eukaryotic cell, at the same time, DNA not encoding the objective gene is reduced to a minimum. The GFP gene is separated from the pEGFP-N3 plasmid, and acts as a reporter gene to construct the Micro-Linear Vector, then both the new vector and the plasmid are transfected to cells, the results are tested by fluorescence microscope and flow cytometry. The results show that the Micro-Linear Vector has a high effective of transfection and safety in 293, 3T3, CNE2 and B95-8 cell lines, at the same time it is less toxicity than the plasmid. We can get the rudiments of conclusion that Micro-Linear Vector has high effection of the transfection and more safety than tradition plasmid in eukaryotic cell.
3T3 Cells ; Animals ; CpG Islands ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Kidney ; cytology ; Mice ; Promoter Regions, Genetic ; genetics ; Transfection ; methods

Objective To analyze the mutation of common driver genes in patients with non-small cell lung cancer(NSCLC)in eastern Henan Province.Methods A retrospective analysis was performed on 661 patients with NSCLC admitted to the First People's Hospital of Shangqiu city from March 2022 to July 2023.Five kinds of gene mutation detection kits(fluorescent PCR)were used for gene detection in all enrolled patients.The relationship between the clinical features and the status of each driver gene was analyzed by statistical methods.Results In the 661 patients with NSCLC,the mutation rates of EGFR,KRAS,ALK,ROS1,PIK3CA,BRAF,HER2,RET,MET14 and NRAS were 47.35%,9.68%,5.45%,1.82%,2.87%,1.82%,1.21%,0.91%,0.61%and 0%.Mutations in EGFR,ROS1 and HER2 were more likely to occur in women(P<0.05),while KRAS mutations were more common in men(P<0.05).The mutation rates of EGFR,KRAS and ALK in adenocarcinoma was significantly higher than that in squamous cell carcinoma and NSCLC.NOS(P<0.05),and the mutation rate of PIK3CA in NSCLC.NOS was the highest.The mutation rate of KRAS gene in stage Ⅰ+Ⅱ was significantly higher than that in stage Ⅲ+Ⅳ(P<0.05),and there was no significant correlation between other genes and clinical stage.Compared with smokers,the mutation rate of total driver gene was significantly higher in non-smokers(P<0.05).EGFR,ALK,PIK3CA,ROS1,BRAF and HER2 were more common in non-smokers(P<0.05),while KRAS gene was more common in smokers(P<0.05).The mutation rate of 10 driver genes in sediment cell block samples was 78.67%,and the detection rate was significantly higher than that in other types of samples(P<0.05).Conclusion Com-mon driver genes such as EGFR,KRAS and ALK are correlated with gender,pathological type,clinical stage and smoking.Qualified samples of sediment cells have obvious advantages for gene detection and could be widely pro-moted in patients.ARMS-PCR combined detection of 10 genes could be used as the preferred gene detection method for newly diagnosed and treated NSCLC patients.

Background: Classical swine fever virus (CSFV), the causative agent of classical swine fever (CFS), is a highly contagious disease that poses a serious threat to Chinese pig populations. Objectives: Many provinces of China, such as Shandong, Henan, Hebei, Heilongjiang, and Liaoning provinces, have reported epidemics of CSFV, while the references to the epidemic of CSFV in Yunnan province are rare. This study examined the epidemic characteristics of the CSFV in Yunnan province. Methods: In this study, 326 tissue samples were collected from different regions in Yunnan province from 2015 to 2021. A reverse transcription-polymerase chain reaction (RT-PCR), sequences analysis, and phylogenetic analysis were performed for the pathogenic detection and analysis of these 326 clinical specimens. Results: Approximately 3.37% (11/326) of specimens tested positive for the CSFV by RTPCR, which is lower than that of other regions of China. Sequence analysis of the partial E2 sequences of eleven CSFV strains showed that they shared 89.0–100.0% nucleotide (nt) and 95.0–100.0% amino acid (aa) homology, respectively. Phylogenetic analysis showed that these novel isolates belonged to the subgenotypes 2.1c and 2.1d, with subgenotype 2.1c being predominant. Conclusions The CSFV was sporadic in China’s Yunnan province from 2015 to 2021. Both 2.1c and 2.1d subgenotypes were found in this region, but 2.1c was dominant.