Objective To investigate whether ultrasound-mediated microbubble destruction can effectively deliver pEGFP-N1 plasmid into human gingival fibroblasts (HGFs).Methods The primary cultured HGFs were divided into 4 groups,that is,plasmid,microbubble+plasmid,ultrasound+plasmid,and ultrasound+microbubble+plasmid groups.pEGFP-N1 plasmid was used to transfect to HGFs as a gene marker with ultrasound-mediated microbubble destruction.The last group was further divided into subgroups to optimize the transfection conditions.After 48 h,phase-contrast fluorescent microscopy was employed to evaluate the expression of green fluorescent protein (GFP).The cell vitality was measured by the MTT assay.Results The transfection efficiency of the ultrasound+microbubble+plasmid group was higher than other experiment groups.Optimal gene expression was found when ultrasound was radiated at 1.5 W/cm2 for 60 s,microbubble was at a concentration of 10% and plasmid was at a concentration of 6.67 ?g/ml,and the transfection efficiency was highest under this condition.Conclusion Under specific conditions,ultrasound mediated microbubble destruction enhances the reporter gene transfection and expression in HGFs.

OBJECTIVE: To study the expression of IL-33 and its receptor ST2 in the nasal mucosa of allergic rhinitis (AR) patients, and to explore the role of IL-33 and ST2 in the pathogenesis of AR. METHOD: Collected 24 cases of nasal septum deviation of patients with AR as AR group,and selected 20 patients with simple septum deviation as normal control. In addition, collected 20 cases diagnosed with AR, who accepted standardized specific immunotherapy(SIT). RESULT: Immunohistochemical results found more positively stained cells of IL-33 and ST2 in AR patients than normal control group, the difference was statistically significant (P < 0.05), Western-Blot detected that IL-33 and ST2 protein level were significantly higher in AR than the normal control group (P < 0.01), PCR detected that the expression of IL-33mRNA and ST2mRNA were increased in AR group compared with the control group, the difference was statistically significant (P < 0.05). While, the serum IL-33 levels in AR were decreased before treatment (72.37 ± 16.18) ng/L than after six months of SIT (47.35 ± 10.59) ng/L,and the difference was statistically significant (P < 0.05). CONCLUSION IL-33 and its receptor ST2 were highly expressed in the nasal mucosa of patients with AR, suggesting that IL-33/ST2 may play an important role in the pathogenesis of allergic rhinitis. Serum levels of IL-33 were significantly decreased after SIT treatment, suggesting that IL-33 may have a positive correlation with the severity of AR.
Case-Control Studies ; Humans ; Immunotherapy ; Interleukin-33 ; metabolism ; Nasal Mucosa ; metabolism ; Nasal Septum ; abnormalities ; Receptors, Interleukin ; metabolism ; Rhinitis, Allergic ; drug therapy ; metabolism ; pathology

OBJECTIVE: To explore the expression of IL-35, Epstein-Barr virus-induced gene 3 (EBI3) mRNA and IL-12A mRNA in peripheral blood of patients with allergic rhinitis and its significance. METHOD: Peripheral blood were collected from patients with allergic rhinitis (46 cases) and healthy human controls(30 cases). The level of IL-35 in serum was measured by enzyme-linked immunosorbent assay. The subunit EB13 and IL-12A of IL-35 mRNA expressions in peripheral blood mononuclear cell(PBMC) were detected by SYBR Green real-time quantitative PCR. RESULT: IL-35 level in AR group (251.22 +/- 46.27) ng/L was significantly lower compared with that in the normal control group (382.17 +/- 25.41) ng/L, (P < 0.01). The mRNA expression level of EBI3 in the patients with allergic rhinitis was significantly lower than that in the normal control group (P < 0.01), AR group EB13 mRNA level was about half of that in the normal control group. But the mRNA expression level of IL-12A in the patients with allergic rhinitis has not significant difference with that in the normal control group (P > 0. 05). CONCLUSION The decrease of IL-35 and EBI3 mRNA in AR,indicated that IL-35 and EBl3 mRNA may play an important role in allergic rhinitis.
Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Interleukin-12 Subunit p35 ; blood ; Interleukins ; blood ; Male ; Middle Aged ; Minor Histocompatibility Antigens ; Rhinitis, Allergic ; blood ; Young Adult

Allergic rhinitis (AR) is a noninfectious inflammatory response in nasal mucosa caused by allergens, which is contacted by a specific individual. The immune imbalance of Th1/Th2 plays an important role in the pathogenesis of AR. In?terleukin (IL)-33, the novel cytokine of IL-1 family, is an important regulatory factor of allergic diseases, autoimmune diseas?es and various inflammatory diseases. IL-33 is a kind of alarm, which is mainly secreted and released by damaged tissues and cells, especially impaired epithelial cells and endothelial cells. IL-33 binding to its receptor ST2L can activate a variety of immune cells to produce Th2 cytokines, precipitating and maintaining Th2 polarization, increasing AR immune inflamma?tion, which is the new target of AR in research and treatment. In this article, we have done a brief overview for the biological functions of IL-33 and its receptor ST2L and the research progress in the AR.

Objective To investigate the effect of different time of hypoxia reoxygenation (HR) on cell autophagy and cell viability.Methods According to the different reoxygenation time,H9C2 cells were divided into five groups:normal control group (group A),hypoxia group (group B),reoxygen 2h group (group C),reoxygen 12h group (group D) and reoxygen 24h group (group E).The expression of LC3 Ⅱ/LC3 Ⅰ in each groups was detected by Western Blot,and the activity of myocardial cells was detected by MlT.Results MlT results showed that the viability of the myocardial cells after hypoxia 2h/reoxygenation 2h was significantly lower than that in the normal control group (P＜0.05).The ratio ofLC3 Ⅱ/LC3 Ⅰ of the cells in group B (hypoxia group),group C (reoxygen 2h group),group D (reoxygen 12h group),group E (reoxygen 24h group) was significantly higher than that of group A (normal control group) (P＜0.05).Among them,the ratio of LC3 Ⅱ/LC3 Ⅰ of the cells in group C and D were significantly higher than that of group B (P＜0.05),paticlularly the group D was the highest (P＜0.05).The activity of myocardial cells in group D was the lowest among all groups.Conclusion Culturing in the three gas incubator can obviously decrease the viability of H9C2cells.Hypoxia can activate autophagy,and autophagy increases significantly after reoxygenation.Autophagy may be activated most obviously at 12 hours and decrease to the level of hypoxia group at 24 hours.Cells can be damaged by hypoxia,and further increase of cellular damage may occur after oxygen inhalation.At 12 hours,the cell viability is the lowest,and the cell viability may be improved at 24h.

Objective To investigate the correlation between sTREM-1 and inflammatory factors expression in critical patients and its effect on severity of disease and clinical prognosis .Methods sTREM-1 ,TNF-α,IL-6 ,IL-10 levels were checked in serum of 54 patients who admitted to intensive care unit (ICU) on the first day and only sTREM-1 was checked again on the third day .At the same time ,APACHEⅡand survival situation in 28 days were recorded .Results sTREM-1 level in critical patients was positive correlated with TNF-α,IL-6 and APACHEⅡ(P<0 .01) .There was positive correlation between APACHE Ⅱ and TNF-α,IL-6 ,but pearson correlation coefficient between sTREM-1 and APACHE Ⅱ was higher than TNF-α,IL-6 .Compared with the survivor group ,the concentrations of serum sTREM-1 was significantly higer in non-survivor group on the first day and the third day after entering ICU(P<0 .05) .Conclusion sTREM-1 level is positive correlated with inflammatory reaction and the severity of disease .It also has prognostic value for outcome in patients with critical illness .

Objective To evaluate the feasibility and safety of vaginal enlarged amputation of cervix to treat patients with cervical cancer of stage Ⅰ a1 and cervical intraepithelial neoplasia grade Ⅲ(CIN Ⅲ)who were unfit for conization surgery.Methods From July 2002 to May 2007,patients with cervical cancer at stage Ⅰ a1,diagnosed by pathology after loop electrosurgical excision procedure(LEEP),large area CIN Ⅲ(the area of lesion≥3/4 on colposcopy),CIN Ⅲ coexisted with vaginal intraepithelial neoplasia (VAIN)in the superior segment of vagina,CIN Ⅱ-Ⅲ recurrence or with residual lesion,positive margin after conization of cervix,who wanted to preserve fertility and(or)corpus uteri were selected to receive vaginal enlarged amputation of cervix.Results Forty-eight eases including 5 with cervical cancer in stage Ⅰ a1,38 with large area CIN Ⅲ(9 with gland involvement),2 with residual lesion and 2 with positive margin after LEEP,1 recurrence after cold knife conization,received the procedure successfully.The median age was 34 years(range 27-40),median operation time was 60 minutes(range 30-100),median blood loss was 40 ml(range 5-300),and median hospital stay was 10 days(range 7-17).After follow-up 1-39 months,no patient had postoperative complications and recurrence,and all patients resumed normal menstrual cycle and sexual life.Condusion Vaginal enlarged amputation of cervix appears to be a safe and feasible procedure for patients with cervical cancer at stage Ⅰ a1 and CIN Ⅲ who are unfit for conization surgery.

Objective:To investigate the infection of spotted fever group Rickettsiae (SFGR) and Rickettsia mooseri ( R.mooseri) of wild rodents in the field of plague foci in Western Yunnan. Methods:The DNA of liver samples of 2 512 wild rodents captured from the plague foci in Lianghe County, Jianchuan County and Yulong County in Western Yunnan from 2015 to 2016 was extracted by magnetic bead method, and the heat shock protein groEL gene primers were used for nested PCR amplification. Gene sequence splicing and Blast homology comparison were performed using DNAStar 7.1 software and GenBank of the National Center for Biotechnology Information (NCBI) of the United States, respectively, and DNAStar 7.1 and MEGA 6.0 softwares were used to construct phylogenetic trees.Results:The wild rodents infected with SFGR were Mus pahari, Rattus steini, Crocidura attenuata and Suncus murinus (one for each), with a total infection rate of 0.16% (4/2 512); no R.mooseri infection was detected. The SFGR infection rates of wild rodents in the plague foci of Lianghe County and Jianchuan County were 0.49% (3/611) and 0.10% (1/1 029), respectively; no SFGR infection was detected in the wild rodents in the plague foci of Yulong County. The homology analysis showed that the homology between SFGR positive samples and reference sequences was 95.45%-100.00%; some of the groEL gene sequences were highly similar among the four positive samples, and the homology was 89.60%-97.40%. Sequence evolution analysis showed that the sequences of three SFGR positive samples from the plague focus in Lianghe County were clustered in the same branch, and the homology reached 94.40%-97.40%; one positive sample sequence from the plague focus in Jianchuan County was clustered in one branch. Conclusion:SFGR infection rate of wild rodents in the field of plague foci in Western Yunnan is low, and no R.mooseri infection is found.

Objective To assess the outcomes of ischemic stroke patients with atrial fibrillation (AF).Methods Six thousand six hundred and ninety-five patients with acute ischemic stroke,admitted to our hospital from May 2005 to December 2013,were recruited consecutively.These patients were divided into combined AF group (n=583) and non-combined AF group (n=6112).The clinical data,including stroke subtypes,stroke severity,risk factors of stroke,NIHSS scores and Barthel index,and outcomes,including mortality,unfavourable prognosis and recurrence at 3,12,and 36 months after stroke were analyzed.Results The prevalence rate of AF in the patients enrolled in this study was 8.7％ (583/6695).There was a higher frequency of AF in female than that in male,with significant difference (45.8％ vs.33.0％,P＜0.05).The patients from combined AF group were older than those without AF.The percentage of severe stroke in AF patients (34.8％) was significantly higher than that in non-combined AF group (8.3％,P＜0.05).The patients with AF were less likely than the patients without AF to have hypertension (63.8％ vs.73.3％),diabetes (24.9％ vs.32.3％),dyslipidemias (26.1％ vs.31.5％),artery stenosis (17.2％ vs.23.4％),current smoking (22.6％ vs.39.0％),and alcohol consumption (7.5 ％ vs.18.9％),with significant differences (P＜0.05).After adjusting age,gender,stroke subtype,and severity,and risk factors,multivariate analysis showed that there was a higher recurrence risk in combined AF group at 3 months after stroke than that in non-combined AF group (P＜0.05);patients with AF had significantly higher mortality,dependency,and recurrence rates at 12 and 36 months after stroke than those without AF (P＜0.05).Conclusion The long-term prognosis of patients with stroke complicated with AF is poor;therefore,normalized anticoagulant therapy should be taken to decrease the recurrence rate and burdens of stroke in China.

Objective:To investigate the seroepidemiological status of Toxoplasma gondii among humans, dogs and cats and its influencing factors in three counties of Yunnan Province, and to assess the risk of dogs and cats transmitting Toxoplasma gondii to humans and causing disease and epidemics. Methods:Three pestis foci of demestic rodent in Mile City, Mang City and Lianghe County in Yunnan Province were selected as the investigation areas, and blood samples of humans, dogs and cats from 16 natural villages were collected. Serum IgG antibodies against Toxoplasma gondii were detected by indirect enzyme-linked immunosorbent assay, and the influencing factors were analyzed by logistic regression. Results:A total of 368 human serum samples, 307 dog serum samples, and 12 cat serum samples were tested. The positive rates of serum IgG antibodies against Toxoplasma gondii in humans, dogs and cats were 54.62% (201/368), 90.88% (279/307), and 91.67% (11/12), respectively. Logistic regression analysis showed that the risk of Toxoplasma gondii infection (serum IgG antibodies positive) in humans of Mang City and Lianghe County were 8.20 times ( AOR = 8.20, 95% CI: 4.38 - 15.36) and 2.22 times ( AOR = 2.22, 95% CI: 1.24 - 3.97) higher than those in Mile City, respectively, and the risk of Toxoplasma gondii infection (serum IgG antibodies positive) in humans in the age group of 30 - < 40 years old decreased by 57% compared to the age group of 30 years old ( AOR = 0.43, 95% CI: 0.19 - 0.98, P < 0.10). The risk of Toxoplasma gondii infection (serum IgG antibodies positive) in dogs of Lianghe County was 89% lower than that in Mile City ( AOR = 0.11, 95% CI: 0.02 - 0.47). The risk of Toxoplasma gondii infection(serum IgG antibodies positive) in dogs aged 2 years old and older was 2.05 times higher than that in dogs aged younger than 2 years old ( AOR = 2.05, 95% CI: 0.91 - 4.64, P < 0.10). Conclusions:The positive rates of serum IgG antibodies against Toxoplasma gondii in humans, dogs and cats in the three counties where the three pestis foci of demestic rodent are located in Yunnan Province is relatively high. The risk of Toxoplasma gondii infection (serum IgG antibodies positive) in humans, dogs is related to the region and age. The risk of dogs and cats transmitting Toxoplasma gondii to humans and other animals is relatively high. It is necessary to strengthen monitoring in key regions, carry out health education, and take corresponding health measures.