Objective. The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V(Glc NAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important.Methods. Wild type of GlcNAcT-V was transfected into COS-7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with site-directed mutagenesis and transfected into COS-7 cells.Results. It was found that after the cells were added tunicamycin(TM, 1 μ g/ml), the activity was 11.7% of the wild type. The activity of the cells with mutating GlcNAcT-V was about 12.0% of the wild type. RT-PCR showed that there was no significant change in mRNA expression among the three groups.Conclusion. The N-glycosylation is important for its activity. Our results suggest that the N-linked carbohydrates on GlcNAcT-V are required for the posttranscriptional activity of the enzyme.

Objective　To study the transcription effects and cleavage activities of rat caspase-3 specific hammerhead ribozyme (Rz107) in both cell-free conditions and BRL-3A cells. Methods　Rat caspase-3 gene fragment was cloned into the pGEM-T EASY vector under the T7 promoter control. The 32　 P-labeled caspase-3 transcript was the target-RNA. Rz107 genes designed against caspase-3 mRNA were cloned into vector p1.5 between 5'-cis-Rz and 3'-cis-Rz. 32　 P-labeled ribozyme transcripts were incubated with target-RNAs at different conditions and autoradiographed after denaturing gel-electrophoresis. Rz107 was electroporated into BRL-3A cells and the Rz107 expression was analyzed by RT-PCR. Results　In cell-free conditions, Rz107 was active at 37℃. The optimal temperature was 50℃. The Km and kcat were 14.13?nmol/L and 2.31*min-1 respectively. Intracellular cleavage efficiency of Rz107 was 37%, as analyzed by RT-PCR. This indicated that the design of Rz107 was correct, and Rz107 had the activity of common enzymes. Conclusions　Rz107 in cell-free conditions possessed perfect specific catalytic cleavage activity, and it can also cleave the target RNA successfully in cells. The results illustrate the feasibility of ribozyme therapy as a potential alternative approach for treating liver disease caused by apoptosis.

A Marfan syndrome child with contractural arachnodactyly deformity of both hands was admitted to our unit in June 2015. The left and right middle fingers and ring fingers of the child received the continuous " Z" skin flap and " V-Y" plasty to reduce the palmar tension on the skin and release the contracture. Due to the improper therapeutic strategy and the underestimation of effect of underlying lesions of Marfan syndrome on the therapeutic strategy, the right middle finger of child was with ischemic necrosis and amputated, and the finger tip was accompanied with distension and paroxysmal neuropathic pain at a specified future date. The skin flaps of the other three surgical fingers were with local necrosis and scar healing. The child was complicated by distal interphalangeal joints stiffness and some serious complications post-discharge. Combined with this case, the similar disorders are not recommended to carry out one-stage operation.

Objective To investigate the role of epigenetic regulations of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) in the development of diabetic retinopathy and the metabolic memory phenomenon after hyperglycemia was terminated.Methods Diabetic rat model was established by intraperitoneal injection of streptozotocin (STZ).Sixty diabetic rats were randomly divided into 3 groups,poor glycemic control group rats were maintained in poor glycemic control for 4 months;semi glycemic control group rats were maintained in poor glycemic control for 2 months,followed by good glycemic control for 2 additional months;good glycemic control group rats were maintained in good glycemic control for 4 months.Twenty normal rats served as control group.The mRNA expression of PGC-1α and superoxide dismutase 2 (SOD2) of retina were measured by real-time PCR;the expression of PGC-1α and manganese superoxide dismutase (MnSOD) protein were measured by Western blot;the situation of DNA methylation in the promotor region of PPARGC1A was measured by bisulfite sequencing.Results The body-weight in the control group was significantly higher than that in the poor glycemic control group,semi glycemic control group and good glycemic control group (all at P =0.000).The blood glucose value in the poor glycemic control group was significantly higher than that in the control group (P =0.000).The expression levels of PGC-1 α mRNA were significantly lower and the expression levels of SOD2 mRNA were significantly higher in the good glycemic control group,semi glycemic control group and poor glycemic control group than those in the control group (all at P＜0.05).The expression levels of PGC-1α and SOD2 mRNA were significantly different between the good glycemic control group and poor glycemic control group (both at P＜0.05).Compared with the control group,the expression levels of PGC-1α and MnSOD protein were decreased in the diabetic model groups,with significant differences between them (all at P＜0.05).The expression level of PGC-1 α protein was significantly higher in the good glycemic control group than that in the poor glycemic control group (P＜0.05).Diabetes increased DNA methylation in the promotor region of PPARGC1A gene of retina.The DNA methylation level was significantly higher in the poor glycemic control group and semi glycemic control group than that in the control group (P =0.008,0.031).No statistical difference was found between the poor glycemic control group and semi glycemic control group (P ＞ 0.05).Conclusions The expressions of PGC-1o mRNA and protein and MnSOD protein in the retina of STZ induced diabetic rats are decreased,the expression of SOD2 mRNA is increased,the expression changes have metabolic memory characteristics.Increased DNA methylation in the promotor region of PPARGC1A when exposed to high glucose may have a role in the regulation of PGC-1 α expression and metabolic memory.

Objective: To explore the influence of three-level collaboration network of pediatric burns in Anhui province on treatment effects of burn children. Methods: The data of medical records of pediatric burn children transferred from Lu′an People′s Hospital and Fuyang People′s Hospital to the First Affiliated Hospital of Anhui Medical University from January 2014 to December 2015 and January 2016 to September 2017 (before and after establishing three-level collaboration network of pediatric burns treatment) were analyzed: percentage of transferred burn children to hospitalized burn children in corresponding period, gender, age, burn degree, treatment method, treatment result, occurrence and treatment result of shock, and operative and non-operative treatment time and cost. Rehabilitation result of burn children transferred back to local hospitals in 2016 and 2017. Data were processed with t test, chi-square test, Mann-Whitney U test, and Fisher′s exact test. Results: (1) Percentage of burn children transferred from January 2014 to December 2015 was 34.3% (291/848) of the total number of hospitalized burn children in the same period of time, which was close to 30.4% (210/691) of burn children transferred from January 2016 to September 2017 (χ²=2.672, P>0.05). (2) Gender, age, burn degree, and treatment method of burn children transferred from the two periods of time were close (χ²=3.382, Z=-1.917, -1.911, χ²=3.133, P>0.05). (3) Cure rates of children with mild, moderate, and severe burns transferred from January 2016 to September 2017 were significantly higher than those of burn children transferred from January 2014 to December 2015 (χ²=11.777, 6.948, 4.310, P<0.05). Cure rates of children with extremely severe burns transferred from the two periods of time were close (χ²=1.181, P>0.05). (4) Children with mild and moderate burns transferred from the two periods of time were with no shock. The incidence of shock of children with severe burns transferred from January 2014 to December 2015 was 6.0% (4/67), and 3 children among them were cured. The incidence of shock of children with severe burns transferred from January 2016 to September 2017 was 3.9% (2/51), and both children were cured. The incidences and cures of shock of children with severe burns transferred from the two periods of time were close (χ²=0.006, P>0.05). Incidence of shock of children with extremely severe burns transferred from January 2014 to December 2015 was 57.1% (32/56), significantly higher than that of burn children transferred from January 2016 to September 2017 [34.5% (10/29), χ²=3.925, P<0.05]. Shock of 25 children with extremely severe burns transferred from January 2014 to December 2015 were cured, and shock of 9 children with extremely severe burns transferred from January 2016 to September 2017 were cured. The cures of shock of children with extremely severe burns transferred from the two periods of time were close ( χ²=0.139, P>0.05). (5) Time of operative treatment of children with moderate, severe, and extremely severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, 2.065, 2.310, P<0.05). Time of operative treatment of children with mild burns transferred from the two periods of time was close (Z=-0.417, P>0.05). Costs of operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were significantly more than those of burn children transferred from January 2016 to September 2017 (Z=-3.324, t=2.167, P<0.05). Costs of operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.627, 0.808, P>0.05). (6)Time of non-operative treatment of children with mild, moderate, and severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, Z=-2.095, t=2.152, P<0.05). Time of non-operative treatment of children with extremely severe burns transferred from the two periods of time was close (t=0.450, P>0.05). Costs of non-operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were obviously higher than those of burn children transferred from January 2016 to September 2017 (Z=-2.164, t=2.040, P<0.05). Costs of non-operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.146, 1.235, P>0.05). (7) Sixty-seven burn children transferred from January 2016 to September 2017 were transferred back to local hospitals for rehabilitation under the guidance of experts of the First Affiliated Hospital of Anhui Medical University, with 25 patients in 2016 and 42 patients in 2017. Effective rehabilitation rates of burn children transferred back to local hospitals for rehabilitation in 2016 and 2017 were both 100%. Conclusions The three-level collaboration network of pediatric burns treatment in Anhui province can effectively increase cure rate of children with mild, moderate, and severe burns, reduce incidence of shock of children with extremely severe burns, shorten time of operative treatment of burn children with moderate, severe, and extremely severe burns, and time of non-operative treatment of children with mild, moderate, and severe burns, reduce treatment costs of children with moderate and severe burns, and improve rehabilitation effectiveness of children transferred from Lu′an People′s Hospital and Fuyang People′s Hospital to the the First Affiliated Hospital of Anhui Medical University.

Objective To investigate the status of human papillomavirus(HPV)infection in women in Li-aocheng area,and to analyze the relationship between HPV genotypes and thin-prep cytologic test(TCT),so as to provide reference for the diagnosis,treatment and vaccination of cervical cancer in this area.Methods The relevant data of 4 968 women who underwent physical examination in the hospital from Januar-y 2020 to April 2023 were retrospectively analyzed.Cervical exfoliated cells were collected for TCT and HPV genotyping,and the distribution of HPV infection and genotype in women of different ages was analyzed.x2 test was used to compare the differences between groups,and multivariate Logistic regression was used to ana-lyze the influencing factors of different HPV types on cervical lesions.Results The overall HPV infection rate was 21.05％,and there was a significant difference in the overall infection rate among different age groups(x2=42.695,P＜0.001).The HPV infection rate of＞49-59 years old group was the highest(26.85％),followed by＞59 years old group.Among the different types of HPV infection,single infection accounted for the highest proportion(15.12％),followed by double infection(4.07％).The top five high-risk genotypes of HPV infection rate were HPV52,16,58,53,66,and HPV16,52,58 were the main high-risk genotypes in HPV single infection.TCT distribution of 1 046 HPV-positive cases:54.97％had no intraepithelial lesion or malig-nant lesion,29.64％atypical squamous cells of undetermined significance,2.58％atypical squamous cells could not exclude high-grade squamous intraepithelial lesion,11.47％low-grade squamous intraepithelial le-sion,and 1.34％high-grade squamous intraepithelial lesion.The infection rates of HPV16,58 and 66 were re-lated to the severity of cervical cytology(x2 trend=23.765,7.807,16.122,P＜0.05).Multivariate Logistic regression analysis showed that HPV16,18,31,33,35,39,51,52,53,56,58,59,66 genotypes were associated with cervical lesions.Conclusion The risk of HPV infection is high in women in this area,especially in mid-dle-aged and elderly women.High-risk HPV types 52,16 and 58 are the most common infection types in this area.HPV16 is closely related to the degree of cervical lesions.

Objective To assess the efficacy and safety ofintravitreal aflibercept injection (IAI) compared with photodynamic therapy (PDT) in the treatment of Chinese patients with predominantly classic subfoveal choroidal neovascularization (CNV) lesions secondary to neovascular age-related macular degeneration (nAMD).Methods A randomized,double-blind,multi-center phase-3 clinical trial lasting for 52weeks (from December 2011 to August 2014).Subjects were randomized in a 3:1 ratio to either IAI group or PDT-to-IAI group.Subjects in the IAI group received 2 mg IAI at baseline and at week 4,8,16,24,32,40,48,with sham injection at week 28,36.Subjects in the PDT-to-IAI group were forced to receive PDT once at baseline and more time at week 12,24 if PDT retreatment conditions were met.Sham injections were given in PDT-to-IAI group at baseline and at week 4,8,16 and 24,followed by 2 mg IAI at week 28,32,36,40,48.The primary outcome of efficacy were the change in mean Best Corrected Visual Acuity (BCVA) from baseline to week 28,and that of week 52.Safety evaluation included the percentage of subjects who suffered treatment emergent adverse events (TEAEs).Results Among the 304 subjects enrolled,there were 228 and 76 cases in IAI group and PDT-to-IAI group respectively.At week 28,the changes of mean BCVA in IAI group,PDT-to-IAI group compared to baseline were +14.0,+3.9 letters,respectively.At week 52,the changes of mean BCVA in two groups were + 15.2,+8.9 letters respectively with the difference of +6.2 letters (95％CI 2.6-9.9,P=0.000 9).At week 52,the mean foveal retinal thickness in the two groups decreased by-189.6,-170.0 μm,respectively.Subjects with the most BCVA increase in IAI group were those aged ＜65,and those with active CNV lesion area ＜50％ of total lesion area.The most common TEAEs in IAI group and PDT-to-IAI group are macular fibrosis [11.8％ (27/228),6.6％ (5/76)] and BCVA decline [6.6％ (15/228),21.1％ (16/76)].There were 3 cases of arterial thromboembolic events defined in the antiplatelet experimental collaboration group,but all were considered unrelated to interventions.Conclusions The efficacy of aflibercept is superior to that of PDT in nAMD patients in China.The therapeutic effect of aflibercept persisted to week 52 in all subjects.The rate of adverse events was consistent with the safety data of aflibercept known before.

Objective:To explore the association between suboptimal health status (SHS) and plasma IgG N-glycans levels among undergraduates in a college in Shandong Province.Methods:A case-control study was conducted from September to November 2017, 100 college students who underwent physical examinations at Weifang University in Shandong, were selected as study participants based on the inclasion and exclusion criteria of the study. According to the criteria of SHS, the participants were divided into an SHS group ( n=50) and a health control group ( n=50). Plasma IgG N-glycosylaton levels were analyzed by means of ultra-high liquid chromatography (UPLC), and 24 glycan peaks were obtained. The Mann-Whitney U-test and binary logistic regression analysis were performed to investigate the association between IgG N-glycans and SHS. P<0.05 was considered statistically significant. Receiver operating characteristic (ROC) curve analyses were used to evaluate the possibility of plasma IgG N-glycans being a biomarker of SHS. Results:The results of univariate and multivariate analysis showed that GP17 was associated with SHS ( P<0.05), and the relative abundance of initial glycan peaks (GP17) was higher in the SHS group compared with the control group. ROC curve analysis showed that the area under the curve (AUC) of the baseline model was 0.826 (95% confidence interval [ CI]: 0.747-0.905, P<0.001); the AUC of the glycan-based model was 0.631 (95% CI: 0.519-0.744, P=0.002), and the AUC of the combined model was 0.848 (95% CI: 0.763-0.912, P<0.001). Compared with the baseline model, the diagnostic efficiency of the combined model revealed a trend of improvement. Conclusions:The SHS of the students in a college in Shandong Province was associated with an IgG N-glycan level of GP17, which was significantly higher than that of the control group.

A young female patient presented with fever, arthralgia, and rash was diagnosed with adults still's disease. When treated with glucocorticoid steroid, the above patient progressed to anuria, sudden, and confusion. After a teamwork involving different departments, the patient was finally diagnosed with atypical hemolytic uremic syndrome (aHUS) and treated with good outcome. aHUS is a rare disease, while Eculizumab is an orphan drug. The diagnosis and treatment of the patient reveals the importance of multidisciplinary team on the diagnosis and treatment of rare and difficult diseases.