Objective: To construct a recombinant adenovirus (abbreviated as ET-M9-PEX) containing MMP-9 signal peptide and noncatalytic carboxyl-terminal hemopexin domain of human MMP-2, and to use the constructed adenovirus as a drug bioreactor in vivo to enhance the expression of an anti-angiogenesis factor for treatment of tumor by a gene therapy strategy. Methods: Adenovirus vector containing M9-PEX gene was constructed by PCR and gene recombination, and was packaged and amplified in L293 cells to obtain ET-M9-PEX recombinant adenovirus with infective ability. The expression and secretion of PEX in ET-M9-PEX-infected cells were detected by Western-blotting and immunofluorescent staining. The inhibitory effect of ET-M9-PEX-conditioned medium on EC cells proliferation was detected by growth curve and its inhibitory effects on angiogenesis and tumor growth were determined by chicken chorioallantoic membrane (CAM) assay in vivo. Results:ET-M9-PEX was successfully constructed and the expression and secretion of PEX in ET-M9-PEX-infected cells were verified. The ET-M9-PEX conditioned medium significantly inhibited the proliferating rate of EC cells. The tumor weights from ET-M9-PEX-infected PG cells in CAM and gradeⅠvessel number were reduced by 57.57%(P

Objective To identify the effect of specialized language training on the speech rehabilitation of total laryngectomy (TL)patients.Methods Twenty-seven TL patients were trained for esophageal speech by specialized speech-language pathologists from October 2005 to December 2006.Then the participants were followed and evaluated their esophageal speech level after one cycle of training program,1 month,3 months,6 months and 12 months after training program.Results The score of participants' esophageal speech was steadily improved during the 12 months after participating the training program.The score of esophageal speech greatly increased in the third month,and this significant improvement was kept until 12 months after the training program.The esophageal speech quality was negatively correlated with age and surgery area of patients.Conclusions Specialized speech-language training showed positive effect on esophageal speech rehabilitation,and esophageal speech training by specialists is worthy of wide application.

Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the generation of human myeloid derived suppressor cells (MDSCs) relied on peripheral blood monocytes,and to establish efficient induction system in vitro of MDSCs.Methods Kidney transplantation recipients between January and March 2017 were included in this study.Purified CD14 + cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days.Phenotypes and immunosuppressive abilities of induced MDSCs (iMDSCs) were investigated with FACS analyses.Inducible nitric oxide synthase (iNOS) mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs.Results A total of 11 recipients were included in this study.HLA-DR expression decreased sharply after the culture with GM-CSF.iMDSCs showed the similar phenotype characteristics with monocytic-MDSCs (M-MDSCs) as well as significant ability to suppress T cells proliferation and cytokines production.iMDSCs expressed higher levels of iNOS than monocytes,and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA (1 mmol/L).The variations of phenotype and suppressive ability were concentrationdependent,and more significant changes could be revealed in the group of 10 μg/L GM-CSF.Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs,and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway.These results may contribute to establish MDSCs induction system,which would provide a basis for the clinical application of MDSCs.

Objective:To explore the mechanism of the cytotoxicity of human NK cells induced by atorvastatin to colon cancer cell lines. Methods:After colon cancer cells (HCT-116,SW-480,Caco-2) were cultured with different concentrations of atorvastatin, CCK-8 assay was used to assess the effect of atorvastatin on growth of colon cancer cells. The amplification of human NK cells was induced by SCGM medium in vitro. Automatic biochemical analyzer was applied to test the cytotoxicity of NK cells to colon cancer cells which cultured with different concentration of atorvastatin. FCM was used to detect the expression rate of MICA/B on the cells. Results:(1) The cultivation of NK cells:The proportion of NK cells attained to 93. 1% from 4. 5% after cultured for 10 days. (2) The effects of atorvastatin on the growth of the colon cancer cells:After cultured with atorvastatin,the inhibition rate of HCT-116 cells was higher than that in control when the density of atorvastatin increased from 5 μmol/L to 40 μmol/L after 48 h and from 1. 25 μmol/L to 40 μmol/L after 96 h ( P<0. 05 ) . Correlation analysis showed that the concentration of atorvastatin and the growth inhibition rate of HCT-116 cells were positively correlated(r[48 h]=0. 13,r[96 h]=0. 22,P<0. 05). (3) The cytotoxicity of NK cells to colon cancer cells effected after atorvastatin: In different atorvastatin concentrations groups,the cytotoxicity of NK cells to three colon cancer cell lines was all higher than that in control ( P<0. 05 ) . The atorvastatin concentration was from 2. 5 μmol/L to 10 μmol/L for HCT-116 cells,from 5 μmol/L to 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 20μmol/L for Caco-2 cells. Among the three cell lines, the cytotoxicity of NK cells to HCT116 was the highest in the same concentration. (4)NK cells by atorvastatin cutting statins 96 h,the concentration of 20 mmol/L and 40 mmol/L inhibition rate was higher than that of control group,more than other groups on NK cell growth without significant effect. ( 5 ) The impact of atorvastatin on MICA/B expression of colon cancer cells: After cultured with different concentrations of atorvastatin,the expression of MICA/B on colon cancer cells was higher than that in control(P<0. 05). The concentration was 2. 5μmol/L and 5μmol/L for HCT-116 cells,10μmol/L and 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 40 μmol/L for Caco-2 cells. Conclusion:Atorvastatin could inhibit the growth of colon cancer cells (HCT-116,SW-480 and Caco-2) in a dose-dependent manner;and it could enhance the cytotoxicity of NK cells to colon cancer cells;it also could promote the expression of MICA/B of colon cancer cells,and improve the immunogenicity of colon cancer cells.

Objective:To compare the efficacy, recurrence rate and safety of a single intravitreal injection of ranibizumab between type 1 prethreshold and threshold retinopathy of prematurity (ROP).Methods:A retrospective cohort study was conducted.A total 100 eyes from 51 ROP patients in Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University from January 2016 to December 2019 were enrolled.Retcam and indirect ophthalmoscope were used to confirm the diagnosis of 39 cases of type 1 prethreshold ROP (70 eyes) and 18 cases of threshold ROP (30 eyes). All the subjects were given a single intravitreal injection of ranibizumab (IVR) 0.25 mg/0.025 ml as the initial treatment within 24-36 hours after diagnosis.Events such as increased intraocular pressure, intraocular bleeding and endophthalmitis were observed and recorded 1 day after IVR.Retcam photography was used to check whether the fundus lesions was subsided 1 week after IVR.Regular follow-up was conducted once every 2-4 weeks until retinal vascularization.Systemic and ocular complications were recorded.The cure rate, the time of retinal vascularization and the recurrence rate of single IVR for type 1 prethreshold disease and threshold disease were compared.This study was approved by the Ethics Committee of Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University.Results:After a single IVR, 86 of 100 eyes (86.00%) were cured, among which 60 of 70 type 1 prethreshold ROP eyes (85.71%) and 26 of 30 threshold ROP eyes (86.67%) were cured, with no significant difference between the two groups ( χ2=0.09, P=0.65). No systemic adverse reactions were observed after a single IVR treatment.The final retinal vascularization time was (11.86±8.60) weeks in the type 1 prethreshold ROP group and (14.45±10.01) weeks in the threshold ROP group, with no statistically significant difference between the two groups ( t=0.88, P=0.22). Recurrence of ROP appeared in 10 eyes after a single IVR, including 4 eyes (5.71%) in the type 1 prethreshold ROP group and 6 eyes (20.00%) in the threshold ROP group, the recurrent rate was significantly different between the two groups ( χ2=4.76, P=0.03). Six recurrent eyes were treated with IVR for the second time, and 4 recurrent eyes were treated with laser photocoagulation.Five eyes presented with preretinal hemorrhage after IVR treatment, and were recovered without intervention. Conclusions:The treatment of type 1 prethreshold ROP and threshold ROP with single IVR in vitreous cavity is safe and effective.The recurrence rate of threshold ROP is higher than type 1 prethreshold ROP.Retreatment can significantly improve the cure rate.

Objective:To compare the protective effect of corneal protectant hydroxypropyl methylcellulose (HPMC) and balanced salt solution (BSS) on corneal epithelium in vitreous body surgery for proliferative diabetic retinopathy (PDR).Methods:A randomized controlled clinical trial was conducted.Fifty eyes of 50 patients with PDR who underwent minimally invasive vitrectomy in Xuzhou First People's Hospital from September 2018 to January 2019 were enrolled and randomly divided into HPMC group and BSS group, with 25 eyes in each group.All eyes underwent three-channel minimally invasive vitrectomy under non-contact wide-angle lens, and 2% HPMC and BSS were applied to keep the ocular surface moist according to grouping during the operation.The combination of cataract surgery, operation time, application times of 2% HPMC or BSS and the grade of corneal epithelial edema before and after operation were recorded.The patients were followed up for 1 month.Corneal fluorescein sodium staining and tear film break-up time (BUT) test were performed before operation and at 1 day, 3 days, 10 days and 1 month after operation.Schirmer Ⅰ test (SⅠt) and ocular surface disease index (OSDI) questionnaire were carried out before operation and at 1 month after operation.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Xuzhou First People's Hospital (No.xyyⅡ[2018]005). Written informed consent was obtained from each patient prior to any medical examination.Results:The average frequency of application of eye drops in the HPMC group was (5.00±1.56) times, which was significantly less than (50.56±17.00) times in the BSS group ( t=-13.071, P<0.01). The postoperative corneal edema score of the HPMC group was 1.0 (1.0, 1.0) point, which was lower than 2.0 (2.0, 2.0) points of BSS group, and the difference was statistically significant ( Z=-4.909, P<0.01). There were statistically significant differences in postoperative corneal fluorescein sodium scores among 1 day, 3 days, 10 days and 1 month in the two groups (HPMC group: χ2=36.040, P<0.01; BSS group: χ2=50.892, P<0.01). The 1-, 3- and 10-day postoperative corneal fluorescein sodium scores in the HPMC group were significantly lower than those in the BSS group (all at P<0.05). One-day, 3-day, 10-day and 1-month postoperative BUT values were (6.15±2.20), (6.95±2.46), (6.16±2.11) and (5.81±2.92) seconds in the HPMC group, respectively, and were (3.89±1.87), (5.32±2.59), (5.01±2.12) and (4.97±2.10) seconds in the BSS group, respectively.There were significant differences in BUT between the two groups at different time points ( Fgroup=5.240, P<0.05; Ftime=2.846, P<0.05). The preoperative and 1-month postoperative SⅠt values in the BSS group were (12.24±5.55)mm and (9.96±4.53)mm, respectively, showing a statistically significant difference ( t=3.863, P=0.001). The postoperative OSDI score in the BSS group was (51.00±12.04) points, which was significantly higherly than the preoperative value of (47.89±10.95) points, and the difference was statistically significant ( t=-2.111, P<0.05). Conclusions:Compared with BSS, 2% HPMC in minimally invasive vitreous surgery has better protective effect on corneal epithelium of PDR, reduces the frequency of eye drop use during the surgery and shortens the repair time of ocular surface tissue after surgery.

Severe asthma is "asthma which requires treatment with high dose inhaled corticosteroids (ICS) plus a second controller (and/or systemic corticosteroids) to prevent it from becoming 'uncontrolled’ or which remains 'uncontrolled’ despite this therapy." The state of control was defined by symptoms, exacerbations and the degree of airflow obstruction. Therefore, for the diagnosis of severe asthma, it is important to have evidence for a diagnosis of asthma with an assessment of its severity, followed by a review of comorbidities, risk factors, triggers and an assessment of whether treatment is commensurate with severity, whether the prescribed treatments have been adhered to and whether inhaled therapy has been properly administered. Phenotyping of severe asthma has been introduced with the definition of a severe eosinophilic asthma phenotype characterized by recurrent exacerbations despite being on high dose ICS and sometimes oral corticosteroids, with a high blood eosinophil count and a raised level of nitric oxide in exhaled breath. This phenotype has been associated with a Type-2 (T2) inflammatory profile with expression of interleukin (IL)-4, IL-5, and IL-13. Molecular phenotyping has also revealed non-T2 inflammatory phenotypes such as Type-1 or Type-17 driven phenotypes. Antibody treatments targeted at the T2 targets such as anti-IL5, anti-IL5Rα, and anti-IL4Rα antibodies are now available for treating severe eosinophilic asthma, in addition to anti-immunoglobulin E antibody for severe allergic asthma. No targeted treatments are currently available for non-T2 inflammatory phenotypes. Long-term azithromycin and bronchial thermoplasty may be considered. The future lies with molecular phenotyping of the airway inflammatory process to refine asthma endotypes for precision medicine.

Objective:To comparatively analyze for the fundus characteristics of acquired immune deficiency syndrome (AIDS) with cytomegalovirus retinitis (CMVR) between first-visit and non-first-visit in ophthalmology.Methods:A retrospective study was performed for 22 patients (41 eyes) diagnosed as CMVR with AIDS by ophthalmology in the Affiliated Municipal Hospital of Xuzhou Medical University from July 2004 to September 2017. The patients were divided into two groups: one with the first-visit in ophthalmology (FVO) and the other with the first-visit in non-ophthalmology (FVNO). All patients underwent visual acuity, intraocular pressure, slit lamp microscope and indirect ophthalmoscope examinations. Thirty-nine eyes of 21 patients with clear refractive media were examined by color fundus photography, of which 5 patients were examined by FFA. Five patients examined by OCT and B-mode ultrasound. The CMVR were characterized as fulminant type or indolent type. All the 22 patients (41 eyes) except 2 patients (4 eyes) accepted highly active antiroviral therapy, and all patients were treated with ganciclovir intravenously. Nine patients (12 eyes) received intravitreal injection of ganciclovir, 7 patients (10 eyes) underwent vitrectomy because of retinal detachment, 6 patients (7 eyes) gave up surgery because of extensive retinal detachment, and the other 12 eyes did not undergo any eye surgery. All patients were followed for 6-58 months (average 23±39 months). The incidences and fundus characteristics of the patients with fulminant or indolent CMVR were compared and analyzed. Between the two groups, the difference of visual acuity and CD4 +T cell count at the first vist and the last follow-up were analyzed by a t-test, and the macular involvement and spread of the two groups were compared by a chi-square test. Results:Fulminant CMVR showed dense yellow-white necrotic lesions along the great vessels with or without satellite lesions, while indolent CMVR showed sparse yellow-white granular lesions with a little bleeding. The concomitant signs included frost-like dendritic vascular sheath, retinal artery occlusion and optic disc edema. Fourteen eyes of 7 patients in FVO group were fulminant, including 2 patients (2 eyes) with retinal artery occlusion and 1 patient (two eyes) with optic disc edema. In FVNO group, there were 27 eyes of 15 patients, including 21 eyes of 13 cases identified as fulminant type and 6 eyes of 4 patients as indolent type. In the fulminant type, there were 4 patients (6 eyes) with frost-like dendritic vascular sheath, 10 patients (12 eyes) with retinal artery occlusion and 4 patients (4 eyes) with optic disc edema. There was no significant difference in initial visual acuity ( t=-1.534, P=0.133), but there was a significant difference in visual acuity at the last follow-up ( t=-3.420, P=0.001). There was no significant difference in CD4 + T cells between the two groups at the first visit ( t=-0.902, P=0.378). The proportions of macular involvement and 3-4 quadrant involvement in FVO group were significantly higher than those in FVNO group ( χ 2=7.552, 7.865; P=0.006, 0.005). Conclusion:For AIDS patients with CMVR, the first-visit in ophthalmology showed more dense necrotic lesions involving macular and a wider range of lesions than the first-visit in non-ophthalmology.

Objective:To compare the clinical effects of minimally invasive vitreous surgery with and without anti-vascular endothelial growth factor (VEGF) drugs for polypoid choroidal vascular disease (PCV) complicated with vitreous hemorrhage.Methods:A cohort study was performed.Thirty-six consecutive cases (36 eyes) with PCV combined with vitreous hemorrhage who underwent 25G minimally invasive vitreous surgery in Xuzhou First People's Hospital from June 2015 to June 2020 were enrolled.According to surgical methods, the patients were divided into pars plana vitrectomy (PPV) group (24 eyes) receiving vitrectomy only and PPV+ anti-VEGF group (12 eyes) receiving vitrectomy first and intravitreal injection of anti-VEGF drugs one week after the operation.All patients were followed up for at least 6 months.The best corrected visual acuity (BCVA) and central retinal thickness (CRT) of the two groups before treatment, 1 month after treatment and at the last follow-up were measured and compared.Postoperative complications such as recurrence of vitreous hemorrhage and macular scar formation were recorded.This study followed the Declaration of Helsinki and was reviewed and approved by the Medical Ethics Committee of Xuzhou First People's Hospital (No.xyyll[2021]014).Written informed consent was obtained from each patient before surgery.Results:Statistically significant differences were found in BCVA between the two groups before and after treatment ( Fgroup=8.552, P=0.006; Ftime=31.775, P<0.001).The BCVA of the two groups at 1 month after operation and at the last follow-up were significantly improved in comparison with before treatment, and the BCVA at the last follow-up was significantly better than that at 1 month after operation (all at P<0.05).One month after operation and at the last follow-up, the BCVA of PPV+ anti-VEGF group was better than that of PPV group, showing statistically significant differences (both at P<0.05).Statistically significant differences were found in CRT between the two groups before and after treatment ( Fgroup=4.797, P=0.041; Ftime=295.764, P<0.001).One month after operation and at the last follow-up, the CRT of both groups was significantly improved in comparison with before treatment, and the CRT was significantly better at the last follow-up than 1 month after operation (both at P<0.05).The postoperative 1-month CRT of PPV+ anti-VEGF treatment group was lower than that of PPV group, with statistically significant difference ( P<0.05).No statistically significant difference was found in CRT between the two groups at the last follow-up ( P>0.05).Elevated intraocular pressure occurred in 2 eyes and rhegmatogenous retinal detachment in 1 eye in PPV group, accounting for 8.33% and 4.17%, respectively.Cataract aggravated in 2 eyes in PPV+ anti-VEGF group.The incidence of vitreous rebleeding in PPV group and PPV+ anti-VEGF group was 16.67%(4/24) and 8.33%(1/12), respectively, with no significant difference ( P=0.646).The incidence of macular scarring in PPV group and PPV+ anti-VEGF group was 4.17%(1/24) and 33.3%(4/12), respectively, showing a statistically significant difference ( P=0.030). Conclusions:Minimally invasive vitreous surgery is a safe and effective way to treat PCV combined with vitreous hemorrhage.It can improve vision, reduce CRT, and the effect is gradually enhanced in the short term.Intravitreal injection of anti-VEGF drugs can enhance the postoperative effect of PPV and present better vision and anatomical structure of retina.