Objective To investigate the effect of the combination therapy of tirofiban and reteplase on endothelial function,coagulation function and plaque inflammation in elderly patients with ST-elevation acute myocardial infarction (STEMI).Methods 100 patients with STEMI were treated with percutaneous transluminal coronary intervention (PCI) from January 2014 to June 2016 in our hospital.39 cases in the control group used conventional oral aspirin,clopidogrel and statins and other treatment.61 cases in the observation group received tirofiban and reteplase on the basis of the control group.The expression of endothelial microparticles (EMP) was detected by flow cytometry (FCM),and the ICAM-1,high sensitivity C-reactive protein (hs-CRP),tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6),endothelin-1(ET-1) were measured by enzyme-linked immunosorbent assay (ELISA).The thrombin time (TT),activated partial thrombin time (APTT),prothrombin time (PT) and other indicators were measured by PUN-2048A coagulation instrument,then statistical analysis was performed.Results The postoperative levels of EMP,ICAM-1 and ET-1 of control group were (693.46±90.72),(768.58±20.46)μg/L and (31.27±8.18)ng/L,which were significantly higher than those in the observation group [(652.36±67.39),(752.37±25.0)μg/L,(28.22±5.05)ng/L],the differences were statistically significant (t=2.41,2.67,2.68,all P<0.05).After operation,the hs-CRP,TNF-α,IL-6 levels in the control group were (4.16±2.35)mg/L,(4.32±2.02)ng/L,(10.59±3.16)ng/mL,which were significantly higher than those in the observation group [(2.22±1.47)mg/L,(2.74±1.52)ng/L,(6.33±2.24)ng/mL],the differences were statistically significant(t=2.65,2.67,3.42,all P<0.05).The postoperative TT,PT,APTT in the observation group were (26.31±3.18)s,(14.34±1.67)s,(27.20±4.12)s,which were significantly longer than those in the control group [(24.03±2.84)s,(12.56±1.43)s,(24.55±3.62)s],the differences were statistically significant(t=2.15,2.31,2.65,all P<0.05).Conclusion Tirofiban combined with reteplase can improve endothelial function,inhibit inflammatory reaction and regulate coagulation function.

Objective To investigate the clinical feature, treatment and prognosis of severe hand-footmouth disease without typical skin lesions. Methods Clinical data from 24 patients with severe hand-footmouth disease without typical skin lesions collected from January 2010 to May 2010 were retrospectively analyzed.Results There were 17 males and 7 females among the 24 patients. Of them, 3 patients with positive EV71 showed no skin lesions at the first visit, 2 presented with only herpes of mouth, 3 with only skin rashes in gluteal regions, 5 with only skin rashes on the palms, 2 with only skin rashes of the knees, 9 with skin rashes in hands, feet and gluteal regions simultaneously. The skin rashes were sparse with an atypical distribution.Fever occurred in all the 24 patients and lasted 6 - 7 days. Nervous system was involved in all the patients,and pulmonary hemorrhage occurred in 4 patients. RT-PCR and real-time PCR showed that the pharyngeal swab was positive for EV71 in 13 patients, for CoxA16 in 2, for other enterovirus in 3, and feces samples were positive for EV71 in 9 patients, for CoxA16 in 1, for other enterovirus in 2. Five patients were positive for EV71 in both pharyngeal swab and feces samples. Four patients died of multiple organ failure, the other 20 patients were cured and discharged from hospital. Conclusions The major pathogen causing severe hand-footmouth disease is EV71 in the 24 patients; children under 2 years are liable to this disease; high fever is common with various rashes. Early recognition of atypical skin lesions and timely management are most important for the control of severe hand-foot-mouth disease and improvement in overall survival of patients with this entity.

Objective To determine the level of interleukin (IL)-10 in sera and culture supernatants of CD4+CD25+T cells from children with atopic dermatitis (AD),and to evaluate its relationship with clinical course and severity of AD.Methods Totally,46 children with AD and 31 healthy controls were included in the study.All the patients were divided into 3 groups,i.e.,mild (n =10),moderate (n =16) and severe (n =20) group,according to severity scoring of atopic dermatitis (SCORAD) score.Venous blood samples were obtained from the patients and healthy controls.CD4+CD25+ regulatory T cells were separated from the blood samples by magnetic cell sorting (MACS) system in two steps and cultured in vitro.Enzyme linked immunosorbent assay (ELISA) was conducted to quantify the level of IL-10 in sera and culture supernatants of CD4+CD25 + T cells from these subjects.Analysis of variance was carried out to compare the level of supematant and serum IL-10 between the patients and controls,and Pearson correlation analysis to assess the relationship between the level of IL-10 and SCORAD score.Results The patients with mild,moderate and severe AD showed a similar serum IL-10 level compared with the healthy controls ((43.10 ± 25.07) pg/ml,(68.40 ± 36.65) pg/ml and (55.55 ± 41.97) pg/ml vs.(58.27 ± 36.84) pg/ml,all P ＞ 0.05).The level of supernatant IL-10 secreted by CD4+CD25+ T cells from the controls was significantly higher than that from the patients with severe AD ((55.15 ± 11.15) pg/ml vs.(27.25 ± 7.01) pg/ml,P ＜ 0.05),but similar to that from the patients with mild and moderate AD ((52.96 ± 11.69) pg/ml and (49.86 ± 9.18) pg/ml,respectively,both P ＞ 0.05).The level of secreted IL-10 was negatively correlated with SCORAD score (r =－0.757,P ＜ 0.01),whereas the serum level of IL-10 showed no statistical correlation with SCORAD score.Conclusion CD4+CD25+ T cells and IL-10 may be implicated in the development of AD.

Objective To assess the associations of anti-Smith antibodies with clinical manifestations and disease activity in children with systemic lupus erythematosus (SLE).Methods According to SLE disease activity index (SLEDAI) score,72 children with SLE were divided into the active group and inactive group.An immunoblotting method was used to detect serum anti-Smith antibodies in these subjects.Chi-square test was conducted to assess the associations of anti-Smith antibodies with clinical manifestations and disease activity in these patients.Results Of these patients,28 (38.9％) were assigned into the inactive group,and 44 (61.1％) to the active group.Anti-Smith antibodies were detected in 17 (23.6％) patients,but not in the other 55 (76.4％) patients.Elevated incidence rate of kidney injury was observed in anti-Smith antibody-positive patients compared with anti-Smith antibody-negative patients (70.6％ (12/17) vs.41.8％ (23/55),P ＜ 0.05).Meanwhile,the positivity rate of anti-Smith antibodies was 31.8％ (14/44) in the active group,significantly higher than that in the inactive group (10.7％,3/28,P ＜ 0.05).Conclusions Anti-Smith antibodies are not only an important indicator for the diagnosis of SLE,but also a risk factor for disease exacerbation and kidney injury in children with SLE.

ObjectiveTo evaluate the roles of imbalance between peripheral blood T helper 17 (Th17) cells and CD4+CD25+ regulatory T(Treg) cells in the pathogenesis of atopic dermatitis (AD).Methods Peripheral blood samples were obtained from 52 patients with AD aged 2-14 years and 30 age- and sex-matched healthy controls.Flow cytometry was performed to detect the percentage of Th17 cells and Treg cells in peripheral blood.Meanwhile,enzyme linked immunosorbent assay(ELISA) was carried out to detect the serumlevel of interleukin (IL)-6 and transforming growth factor (TGF)-β1.Results The children with AD showed a higher percentage of Th17 cells but a lower percentage of Treg cells in CD3+ T cells compared with the controls (( 1.20 ± 0.41 )％ vs.(0.54 ± 0.28)％,t =2.58,P ＜ 0.05; (2.29 ± 0.67)％ vs.(5.95 ± 0.45)％,t =15.23,P ＜ 0.01 ).Moreover,the serum level of IL-6 was significantly higher,while that of TGF-β1 was lower in patients with AD than in the controls ((5.12 ± 0.45) ng/L vs.(3.89 ± 0.38) ng/L,t =2.59,P＜ 0.05; (57.65 ± 10.78) ng/L vs. (81.18 ± 7.78) ng/L,t =5.41,P ＜ 0.01 ).ConclusionsChildren with AD experience a change in the percentage of Thl7 cells and Treg cells in peripheral blood as well as in the serum level of IL-6 and TGF-β1,and the imbalance between Th17 cells and Treg cells in peripheral blood may contribute to the development of AD.

Objective To investigate the differences in clinical efficacy and safety between thrombolysis followed PCI (percutaneous coronary intervention) and primary PCI in patients with acute STEMI (ST elevation myocardial infarction). Methods A total of 215 STEMI patients who visit our clinic within 12 h since onset of their symptoms from May 2013 to January 2015 were enrolled. All eligible patients were divided into Early PCI group(n=68) and pPCI group (n=147) based on whether or not they received injection of recombinant human prourokinase thrombolytic therapy before their visit. Immediate TIMI (Thrombolysis In Myocardial Infarction) flow grade of infarct-related artery (IRA) before and after PCI treatment, post?operative CTFC (Corrected TIMI Frame Count) and TMPG (TIMI myocardial perfusion grade) were compared between these two groups. The incidence of bleeding during hospital stay , left ventricular function at 6 month after intervention and major adverse cardiac events (MACE) were all observed. Rusults There is no obvious difference between the baseline of two groups. Before PCI, the proportion of TIMI grade 2-3 was higher in Early PCI group (77.9%vs 20.4%,P<0.05)than that in pPCI group;but there was no significant difference in the proportion of TIMI grade 2-3 between these two groups after PCI (P>0.05). CTFC and peak value of serum CK-MB were lower [(27.7 ± 5.0) vs (32.6 ± 7.1), P<0.05;(225.8 ± 108.3) U/L vs (283.4 ± 110.6) U/L, P<0.05] and rate of TMPG 3 is higher (82.4%vs 68.7%, P<0.05)in Early PCI group than those in pPCI group. No significant difference was found in the incidence of bleeding and MACE during hospital stay and Left ventric?ular function at 6 months after operation between these two groups. By contrast, LVEFs were higher while LVEDds (LVED diameter) were lower after 3 and 6 months of the intervention compared to those before intervention in both groups (P <0.05). Conclusion It is a safe and effective reperfusion strategy for STEMI patients to receive rhPro-UK thrombolytic thera?py followed early PCI as an alternative way to those who failed to receive pPCI on time. It didn′t increase the occurrence of bleeding complications and MACE, and at the same time it presented the same benefit in improving recent cardiac function as pPCI did.

Objective: To investigate the seroprevalence and influencing factors of serum neutralizing antibodies among SARS-CoV-2 infected individuals, so as to provide the evidence for developing the health management and COVID-19 vaccination strategy among SARS-CoV-2 infected individuals. Methods: Recovered SARS-CoV-2 infected individuals from January 1st, 2020 to February 10th, 2021 in Zhejiang Province were recruited in March 2021. Participants' demographics, underlying diseases, date of definitive diagnosis and severity of clinical symptoms were collected using questionnaire surveys, and serum neutralizing antibody against SARS-CoV-2 was detected using a fluorescent immunoassay. In addition, factors affecting the seropositivity of neutralizing antibody against SARS-CoV-2 were identified using a multivariable logistic regression model. Results: A total of 559 SARS-CoV-2 infected individuals were enrolled, including 480 confirmed cases and 79 asymptomatic carriers, with an median (interquartile range) age of 47.00 (22.00) years, and all participants had never received COVID-19 vaccination. The median (interquartile range) duration from diagnosis to serum sampling was 387.00 (11.00) days, and the seroprevalence of neutralizing antibody against SARS-CoV-2 was 83.90%. The serum neutralizing antibody against SARS-CoV-2 was all positive 9 months after diagnosis, and the seroprevalence of neutralizing antibody against SARS-CoV-2 appeared no tendency towards a decline with time within 14 months after diagnosis (P>0.05). Multivariable logistic regression analysis showed that women were 1.892 times (95%CI: 1.169-3.064) more likely to produce serum neutralizing antibodies against SARS-CoV-2 than men, and mild, common and severe/critically ill SARS-CoV-2 infected cases were 2.438 (95%CI: 1.305-4.557), 4.481 (95%CI: 2.318-8.663), and 23.525 (95%CI: 2.990-185.068) times more likely to produce serum neutralizing antibodies against SARS-CoV-2 than asymptomatic carrier, respectively. Conclusions The seroprevalence of neutralizing antibody was 100.00% among SARS-CoV-2 infected individuals within 9 months after diagnosis. Individuals' gender and severity of clinical symptoms correlate with the seroprevalence of neutralizing antibody against SARS-CoV-2.

Objective: To investigate the accuracy and repeatability of contrast-enhanced transthoracic echocardiography for measurements of right ventricular structure and function. Methods: The apical four-chamber views and the three-dimensional full-volume images of the right heart were collected from 12 beagles with unenhanced and contrast-enhanced transthoracic echocardiography. The intimal display rate of the right ventricular segments, right ventricular end diastolic longitudinal dimension (RVLD), right ventricular end diastolic area (RVEDA), right ventricular end systolic area (RVESA) and right ventricular fractional area change (RVFAC) were evaluated respectively with two-dimensional unenhanced and contrast-enhanced echocardiography. Right ventricular three-dimensional full-volume images were processed and analyzed by TomTec software, and right ventricular end diastolic volume (RVEDV), right ventricular end systolic volume (RVESV) and right ventricular ejection fraction (RVEF) were measured respectively with three-dimensional unenhanced and contrast-enhanced echocardiography. The measurements of pathological specimen were taken as the gold standard, the accuracies of measuring RVEDVand RVLD by different methods were evaluated. All indexes were measured repeatedly by the same observer and different observers to assess the intraobserver and interobserver reproducibilities of different methods. Results: ①The intimal display rate of the right ventricular segments was higher with contrast-enhanced echocardiography than that with unenhanced echocardiography (P<0.05). ②The measurements of RVEDV by three-dimensional contrast-enhanced echocardiography correlated well with the measurements by anatomical specimens. And the correlation was higher (0.916 vs 0.843), the consistency was better than that by unenhanced echocardiography. The measurements of RVLD by two-dimensional contrast-enhanced echocardiography correlated well with the measurements by anatomical specimens. And the correlation was higher (0.928 vs 0.850), the consistency was better than that by unenhanced echocardiography. ③For inter- and intraobservers reproducibilities, the interclass correlation coefficients of RVLD, RVEDV, RVESV, RVEF, RVEDA, RVESA, RVFAC with contrast-enhanced echocardiography were higher and 95% confidence interval ranges were smaller than those with unenhanced echocardiography. Conclusions Contrast-enhanced transthoracic echocardiography can improve the accuracy and repeatability for measurements of right ventricular structure and function, providing a new evaluation method for patients with poor image quality of the right ventricle in clinical practice.

Finding stable expression sites on the chromosomes of Chinese hamster ovary (CHO) cells is an effective method to solve the problem of unstable expression of CHO cells in long-term culture. Our group used lentiviral transfection to integrate the tracer gene (Zsgreen1) into the chromosome of CHO cells and found multiple potential stable expression sites. This study verified the ability of one of the sites located in the 148052-148157 bp region on chromosome NW_003614241.1 to stably express exogenous proteins.The expression of Zsgreen1 gene was first observed, and CRISPR/Cas9 technology was then used to integrate the enhanced green fluorescent protein (EGFP) gene into this site. Three strains of EGFP gene integrated cells were obtained. After 60 generations of suspension culture, the fluorescence intensity of the cells had no significant changes, which proved that this site can stably express the EGFP gene. The same method was used to construct recombinant CHO cell lines expressing the human serum albumin (HSA) gene, and was verified by Western blot that this site could express and secrete HSA. It shows that the above-mentioned sites can be integrated and can stably express exogenous proteins.

Despite its more than 100-year history in experimental and clinical use, photodynamic therapy (PDT) is only starting to be appreciated for its full potential. PDT combines a photosensitizer and light in the presence of oxygen to treat cancer and other disorders. This paper reviews the molecular mechanism of PDT at the cellular level as well as in therapeutic settings in vivo. The availability of multiple photosensitizers with different structures and functional properties makes PDT an extremely versatile and, conversely, a challenging approach to cancer therapy. The advancing understanding of molecular pathways helps to design improved regimens. As most cancers are being treated with combined therapies, PDT is being integrated into rationally designed regimens that exploit molecular responses to PDT for improved efficacy.
Humans ; Neoplasms ; drug therapy ; Photochemotherapy ; Photosensitizing Agents ; therapeutic use