Objective:To express fusion protein of mouse thymic stromal lymphopoietin (TSLP) and HIV-1 gp120BAL V1/V2 subdomain in 293F cell.Methods:Full length of the V1V2 sequence from BAL isolate was fused with the C-terminus of mouse thymic stromal lymphopoietin (TSLP) and sub-cloned into pCEP-Pu vector to construct the eukaryotic expression plasmid-pCTV1V2BAL.The recombinant plasmid was confirmed by enzyme digestion and sequencing , then transfected into 293 F cells using PEI as a transfection reagent .The fusion protein was purified by metal chelate affinity chromatography and characterized by SDS -PAGE and Western blot . The epitopes of V1/V2 in fusion protein were identified by ELISA .Results:The SDS-PAGE and Western blot results showed that there were highly heterogeneous glycoprotein bands at the site between 35 kD and 55 kD, which reacted with anti-mTSLP rabbit polyclonal antibody and anti-His tag mouse monoclonal antibody .The ELISA analysis showed that antibodies to V 1/V2BAL existed in the sera of HIV-1 positive patients.Conclusion:The mTSLP-V1/V2 fusion protein was successfully expressed in 293F cells, which may be useful for HIV-1 vaccine research .

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.

Objective:To observe the clinical features, treatment and prognosis of ciliary body tumors.Methods:A retrospective clinical study. From November 2011 to March 2023, 8 cases (8 eyes) with ciliary body tumours confirmed by pathohistological examination at the Department of Ocular Oncology, Beijing Tongren Hospital were included in the study. Patients' age, gender, involved eyes, symptoms, best corrected visual acuity (BCVA), intraocular pressure, cataract, lens subluxation, and imaging manifestations were collected in detail. All affected eyes were treated surgically. The follow-up time after surgery ranged from 1 to 10 years. The patients' clinical presentation as well as imaging, pathohistological features and treatment and prognosis were analysed retrospectively.Results:Among 8 cases (8 eyes), there were 3 males (3 eyes) and 5 females (5 eyes), 3 and 5 eyes in the right and left eyes, respectively. The median age was 44 years. Ciliary body medulloepitheliomas, melanoma, squamous cell carcinoma, leiomyoma, schwannoma, and adenoma of the nonpigmcnted ciliary epithelium were in 2, 2, 1, 1, 1, and 1 eyes, respectively. All reported decreased or loss of vision. Cataract, vitreous opacity, red eye and or (ocular pain), retinal detachment, lens subluxation, and secondary glaucoma were 6, 4, 4, 2, 1, and 1 eyes, respectively. Diagnostic imaging was consistent with pathological findings in 3 eyes. The first surgery was performed for enucleation and orbital implantation in 2 eyes, the patients were 9 and 10 years old with medullary epithelioma; the follow-up time after surgery was 1 and 5 years, respectively. Local tumour resection was performed in 6 eyes. Among them, 3 eyes with benign tumours were followed up for 1 to 9 years after surgery; 2 eyes showed significant improvement in visual acuity, 1 eye with adenoma of the nonpigmcnted ciliary epithelium had a preoperative BCVA of finger count/1 m, and a postoperative BCVA of 0.5, and 1 eye with leiomyoma had a preoperative BCVA of 0.06, and a postoperative BCVA of 0.5; and 1 eye was lost to follow-up. Malignant tumour in 3 eyes, of which 2 eyes recurred after surgery. Re-operation for enucleation and local tumour excision combined with local cryotherapy in 2 eyes of recurrence were 1 eye each, respectively. The follow-up period after surgery was 2 and 4 years, respectively. No recurrence after surgery in 1 eye, but there was no significant improvement in visual acuity during follow-up. No recurrence or metastasis was observed in any of the eyes during the follow-up period or at the final follow-up.Conclusions:Ciliary body tumour types and clinical presentations are complex and varied; imaging can detect tumours but is poor at determining the nature of the lesion. Benign tumours do well with local excision surgery; malignant tumours do well with enucleation.

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.

Objective To observe the thickness changes in the peripapillary retinal nerve fiber layer(RNFL)and the ganglion cell-inner plexiform layer(GC-IPL)in the macular region,so as to analyze the pathological rules of retinal nerve injury in diabetes retinopathy(DR)patients and explore the related risk factors.Methods A total of 164 patients(164 eyes)with type 2 diabetes who visited the Department of Ophthalmology and Department of Endocrinology of Shaanxi Pro-vincial People's Hospital from January 1,2017 to January 1,2018 were selected as the subjects.According to the condition of concomitant DR,these subjects were divided into three groups:non-DR(NDR)group(56 eyes of 56 patients),mild non-proliferative DR(NPDR)group(53 eyes of 53 patients)and moderate NPDR group(55 eyes of 55 patients).Healthy individuals who underwent physical examinations in the same period were selected as the control group(50 eyes of 50 pa-tients).Optical coherence tomography and optical coherence tomography angiography were used to measure the peripapil-lary RNFL,macular GC-IPL,and macular vessel density(VD)of all subjects.Comparison among groups and correlation analysis were conducted on the above retinal nerve injury indicators.Results Compared with the control group and NDR group,the average,upper and lower peripapillary RNFL in the mild NPDR group and moderate NPDR group were signifi-cantly thinner,and the differences were statistically significant(all P＜0.05).The differences in macular GC-IPL thickness at the average,upper,upper temporal,lower temporal,lower,lower nasal,and upper nasal regions in all groups were sta-tistically significant(all P＜0.05);pairwise comparisons showed that compared with the control group,the GC-IPL thick-ness in each quadrant of the macular area of patients in the NDR group,mild NPDR group and moderate NPDR group de-creased significantly(all P＜0.05),with the upper GC-IPL thinning being the most significant;compared with the NDR group,the GC-IPL thickness in the upper,average,and upper temporal macular area of patients with mild NPDR decreased significantly(all P＜0.05);compared with the mild NPDR group,the GC-IPL thickness in the upper macular area was thin-ner in the moderate NPDR group,and the difference was statistically significant(P＜0.05).There were statistically signifi-cant differences in the average and inner ring VDs in the macular area among the groups(both P＜0.05).The correlation analysis results showed that there was a significantly positive correlation between the peripapillary RNFL thickness and the average VD in the macular area(r=0.517,P＜0.01),while the average GC-IPL thickness in the macular area was weakly positively correlated with the average VD in the macular area(r=0.279,P＜0.01).Conclusion The peripapillary RNFL thickness objectively reflects the degree of retinal neuron injury,and is a key index to evaluate the degree of retinal nerve injury in diabetes patients.

Objective To investigate the effects of microRNA-26a-5p(miR-26a-5p)on high glucose-induced retina Müller activation and apoptosis by regulating PTEN/PI3K/Akt signaling pathway,and the potential mechanism of diabetic retinal neurodegeneration.Methods Various concentrations of high glucose were added into rMC-1 culture.CCK-8 and flow cytometry was used to examine cell proliferation and apoptosis respectively.The regulatory effects of miR-26a-5p on the expressions of PTEN,PI3K and Akt were observed by real-time PCR;the expression levels of IL-1β and IL-6 in Müller cells were examined by ELISA.The data were processed by Graphpad 8.0 software.Results Müller cells grew actively in high-glucose stimulation culture.Compared with the control group,the activity of Müller cells stimulated by 50 mmol/L glucose increased gradually at 12 h and 24 h,but decreased at 48 h after stimulation,when Müller cells apoptosis increased.The difference between the groups was statistically significant(P＜0.05).The expression of miRNA-26a-5p decreased,that of PTEN increased,and those of PI3K and Akt decreased.Meanwhile,IL-1β and IL-6 levels were significantly increased in Müller cells.miR-26a-5p over-expression alleviated injuries to high glucose stimulated retinal Müller cells by inhibiting PTEN,which upregulated the expression of PI3K/Akt and downregulation of IL-1β and IL-6(P＜0.01).Conclusion Upregulating miR-26a-5p protects Müller cells against apoptosis,probably through regulation of PTEN/PI3K/Akt and affecting the production of inflammatory factors.

ObjectiveTo explore the status of mild cognitive impairment （MCI） among the elderly in Shanghai communities， and to identify the factors associated with MCI. MethodsThe Montreal Cognitive Assessment-Basic （MoCA-B） was used to screen for MCI in elderly adults. Logistic regression analysis was conducted to identify the factors associated with MCI. ResultsAmong 629 participants， 226 （35.90%） were positive for MCI. Older age （OR=1.04， 95%CI： 1.01-1.07）， lower family income （average OR=2.20， 95%CI： 1.01-4.80； poor OR=2.59， 95%CI： 1.03-6.50）， hearing impairment affecting daily life （OR=1.86， 95%CI： 1.19-2.91）， and anxiety symptoms （OR=1.58， 95%CI： 1.02-2.44） were associated with the positive for MCI. Living in central urban area （OR=0.57， 95%CI： 0.37-0.89） and having higher social support （OR=0.97， 95%CI： 0.94-1.00） were protective factors for MCI. ConclusionThe current status of MCI among the elderly in Shanghai communities is not optimistic. It is important and necessary to establish the cognitive-friendly community for the elderly.

AIM: To study the protective effect of fenofibrate on diabetic retinal neurodegeneration and observe its effect on miR-26a-5p and its target gene PTEN in the retinal of diabetic mice.METHODS: Diabetic mice models were established and they were gavaged by fenofibrate. H& E staining and transmission electron microscopy were used to observe the impairments of retinal neurons. Real-time PCR was used to examine the expression of miR-26a-5p, and Western blotting was employed to measure the expression of phosphatase and tensin homologue(PTEN)in the retina of diabetic mice. The expression level of nuclear factor-κB(NF-κB), interleukin-1β(IL-1β)and the morphology of neural tissues were observed.RESULTS: When compared with the diabetic mice, fenofibrate significantly attenuated the damage to retinal ganglion cells and the atrophy of retinal nerve fiber layer. While the level of miR-26a-5p was increased and the levels of PTEN and inflammatory mediators were significantly decreased in the retina of fenofibrate treated diabetic mice, with significant statistical significance(P<0.05).CONCLUSIONS: Fenofibrate protects against diabetic retinal neurodegeneration by upregulating miR-26a-5p and inhibiting PTEN, attenuating the inflammatory response and alleviating retinal cell injury.