Objective To observe the effects of PTD-hepatitis B core antigen (HBcAg) induced murine bone marrow-derived dendritic cells (DCs) maturation on T lymphocyte proliferation in vitro.Methods Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granu|ocyte-macrophage colony-stimulating factor (rGM-CSF) and recombinant interleutin-4 (rIL-4)for 5 days.Tumor necrosis factor (TNF)-a,HBcAg and PTD-HBcAg were added to induce DCs maturation.The distribution and localization of intracellular immunofluorescence were observed by confocal microscopy,and DCs phenotypes were analyzed by flow cytometry.The level of IL-12 p70 in the supernatant was detected by enzyme linked immunosorhent assay (ELISA).The proliferation of T lymphocytes was performed by using cell counting kit-8 (CCK-8).All data were analyzed using t test.Results DCs were cultured and identified successfully.Recombinant PTD-HBcAg could penetrate into DCs cytoplasm while recombinant HBcAg was detected on the surface of cells.DCs surface molecules,such as CD80,CD86 and major histocompability complex (MHC) II were upregulated by PTDHBcAg;IL-12 p70 levels induced by 50 mg/L and 100 mg/L recombinant PTD-HBcAg were (142.50±18.31) ng/L and (124.30±15.12) ng/L,respectively,which were significantly higher than those induced by recombinant HBcAg [(42.31±4.21 ) ng/L,t = 9.234 and 9.045,respectively,P＜0.05].The proliferation of T lymphocytes induced by PTD-HBcAg was much higher than that in HBcAg group or positive control TNF-a group.Conclusions PTD-HBcAg could penetrate membrane of DCs and promote the differentiation and maturation of DCs.PTD-HBcAg could up-regulate the expressions of costimulatory molecules on cell surface of DCs,and enhance the ability of DCs on stimulating T lymphocytes proliferation and IL-12 p70 production.

Objective To investigate the effect of hepatitis B virus core protein (HBc) dimer interfaces amino acids mutation on nucleocapsid assembly and HBV DNA replication. Methods Based on HBc three dimension structure, four HBc dimer interfaces domain mutation plasmids, pHBc14-18M,pHBc120-135M,pHBc23-39M and pHBc122-139M were constructed in pcDNA3.1 vector by PCR site-directed mutagenesis, there was a flag-tag at the C-terminal of all mutants for easy detection. Wild type core protein plasmid 1-183flag was also constructed as a positive control. The 4 mutants were cotransfected HepG2 cells with pHBV1.2 core negative plasmid (pHBV1.2-core-) ,which contained 1.2 copies of HBV whole genome but the core protein would not express due to a stop codon. The capsid formation, HBV pregenome(pgRNA) and HBV DNA replication mediate were analyzed by native agarose gel electrophoresis and Western blot, Northern blot and Southern blot , respectively. The 4 mutants were also cotransfected HepG2 cells with HBV wild type plasmid pHBV1.2 and examined by Southern blot. Virions in the medium were determined by native agarose gel electrophoresis and Western blot. Results Cotransfecting HepG2 cells with pHBV1.2-core- plasmid, pHBc14-18M,pHBc120-135M and pHBc122-139M mutant groups formed nucleocapsid-like structure but pHBc23-39M could not, Northern and Southern blot displayed no signal in all mutants except 1-183flag conrol group. In pHBV1.2 cotransfection experiment, HBV DNA replication was blocked in pHBc14-18M, pHBc120-135M and pHBc122-139M mutant groups, sharply decreased in pHBc120-135M and pHBc122-139M groups, correspondingly virons production in medium were also inhibited. pHBc23-39M mutant exerted no influence on HBV replication. Conclusion pHBc23-39M mutant can neither form nucleocapsid-like structure nor interact with wild type HBc dimmer to interfere HBV replication.On the contrast, pHBc14-18M, pHBc120-135M and pHBc122-139M mutants can form nucleocapsid-like structure by themselves, but this structure does not support HBV DNA synthesis. Besides, they can effectively inhibit wild type HBV DNA replication by contacting with wild HBc dimmers resulting in nucleocapsid dysfunction.

A total of 209 type 2 diabetic patients were divided into three groups:without,with mild to moderate,or severe diabetic lower extremity arterial disease based on the ankle brachial index value.60 healthy subjects were used as a control group.Retinol binding protein4 (RBP4),cystatin C,and biochemical parameters were determined in all subjects.The results showed that RBP4 and cystatin C levels were progressively raised with the increasing severity of lower extremity arterial disease in various groups,being significantly higher in patients with type 2 diabetes mellitus with mild to moderate and severe arterial disease compared with control group (P＜0.05 or P＜0.01).RBP4 and cystatin C levels in groups with lower extremity arterial disease were significantly higher than those in the group without arterial disease (P＜0.01).Pearson correlation analysis showed that RBP4 level was positively correlated with total cholesterol,triglyceride,low density lipoprotein-cholesterol,fasting serum insulin,body mass index,cystatin C etc while negatively correlated with high density lipoprotein-cholesterol (P＜0.05 or P＜0.01).Logistic regression analysis showed that serum RBP4 and cystatin C were significantly associated with lower extremity arterial disease in type 2 diabetes mellitus.

Objective To evaluate the outcomes of penoplasty with penile frenulum lengthening for concealed penis in children.Methods From March 2014 to March 2016,a total of 233 patients with concealed penis who enderwent penoplasty with penile frenulum lengthening were enrolled.The everage age at surgery was 3.7 years (1 year and seven months-12 years).There were 73 cases with obvious small urinary stream,dysuria,or prepuce dilatation when urinating;41 cases with the history of recurrent infections of prepuces;and 9 cases with the history of urinary tract infections.During operation,incise the back side center of the outer plate of the prepuce and fully release the ring-type funicular tissue between the inner and outer plates of the prepuce to make the inner plate fully swell out.After the prepuce is upturned,cut the penile frenulum at the coronary sulcus.Make a V-shaped cut on the left and right sides of the far end of the outer plate cut of the prepuce from the cut of the penile frenulum.Lengthen penile frenulum after the prepuce is pressed off.Cover the dorsal side of the penis with the inner plate of the prepuce and cover its ventral side with its outer plate.Results Mean surgical time for patients was 38 min (30-55 min).All operations were completed successfully and the post-operation follow-up lasts 3 months to 2 years.For all cases,the appearance of the penis is improved.The penis stretches out,the balanus is exposed,the prepuce has no obvious swelling and the scrotum angle of the penis is obvious.No phimosis relapses and there is no obvious scar hyperplasia.Conclusions This surgical procedure is an effective treatment choice for concealed penis.It provides a good cosmetic result.

Objective To study the nucleotide and amino acid sequences of norovirus G Ⅱ.4/Sydney 2012 variants,in China.Methods Twenty-two stool specimens,confirmed as G Ⅱ.4/Sydney 2012-positive were collected from Beijing in the winter of 2012-2013.RT-PCR was performed to target the complete capsid gene.G Ⅱ.4/Sydney 2012 strains from other regions in China were searched and obtained from the GenBank.Nucleotide and amino acid sequences of G Ⅱ.4/Sydney 2012 strains were analyzed,using the CLUSTAL X (Version 1.83)and followed by phylogenetic analysis using Mega version 5.1.Results The complete major capsid nucleotide sequences of thirty-eight G Ⅱ.4/Sydney 2012 strains from seven regions in China were obtained.The VP1 nucleotide and amino acid sequences diversity were 0.1％-3.3％ and 0-3.1％,respectively.Result from phylogenetic analysis demonstrated that the G Ⅱ.4/Sydney 2012 variant shared a common ancestor with both the dominant norovirus G Ⅱ.4 variants Apeldoom 2008 and the New Orleans 2009.G Ⅱ.4/Sydney 2012 variants appeared to have had two A/D/E site combinations at the amino acid level,TSRN-GTT-SNT and TSRN-STT-SNT.Conclusion G Ⅱ.4/Sydney 2012 variant had been circulating in many regions in China.There seemed a high nucleotide and amino acid identity among G Ⅱ.4/Sydney 2012 strains collected from China.G Ⅱ.4/Sydney 2012 variants showed different A/D/E site combination from other pandemic G Ⅱ.4 variants.

Objective To evaluate the long-term efficacy of cervical artificial disc replacement for patients with degenerative cervical canal stenosis.Methods All of 43 patienta underwent single level Bryan cervical artificial disc replacement for degenerative cervical canal stenosis were retrospectively analyzed with a minimum 10 years follow-up in our hospital between December 2003 and December 2007.There were 28 males and 15 females with an average age of 56.7 ± 8.0 years which ranged from 37 to 76 including 1 case of C3.4 level,11 xases of ~ level,26 cases of C~ level and 5 cases of C6,7 level.According to the symptoms and location of compression on radiograph,the patients were divided into 3 groups:15 patients in radiculopathy group,10 patients in myelopathy group and 18 patients in myeloradiculo p~hy group.Radidogical evaluation indexes include global range of motion (ROM),segmental ROM,segmental Cobb angle.Clinical evaluation indexes including Japanese Orthopaedic Association (JOA) score,neck disability index (NDI) and Odom's criteria.The variable data were analyzed by one-way ANOVA,the grading data were analyzed by Kruskal-Wallis H test.The preoperative and postoperative data were analyzed by paired t test.Results At the last follow-up,the Cobb angle of the operative segment was 1.29°±1.34°in myelopathy group(t=4.606,P=0.001),0.71°±2.20°in radiculopathy group (t=2.355,P=0.034),and 0.69°±2.12° in myeloradiculopathy group (t=3.312,P=0.004),which was significantly lower than that before operation.At the last follow-up,ROM of the operative segment in myelopathy group was 6.41 o ±4.87°,which was significantly lower than that before operation (11.46° ±5.19°,t=3.589,P＜ 0.05),and there was no significant difference in the other two groups.There was no significant difference in other imaging parameters among the three groups.JOA scores at the last follow-up were 16.33±0.75 in radiculopathy group (t=5.857,P＜ 0.001),16.00±1.05 in myelopathy group(t=8.337,P＜ 0.001) and 14.78±1.69 in radiculopathy group (t=4.045,P＜ 0.001);NDI were 13.07％±5.90％ in radiculopathy group (t=7.097,P＜ 0.001),12.60％±4.22％ in myelopathy group (t=7.319,P＜ 0.001) and 23.11％±14.18％ in radiculopathy group (t=4.229,P＜ 0.001),which were all significantly improved than those before operation.The excellent and good rate of Odom's criteria (H=2.719,2.411,P＜ 0.05),JOA (LAD-t=3.770,4.080,P＜0,05) and NDI(LAD-t=2.850,2.643,P＜ 0.05) at the last follow-up of the patients in radiculopathy group and myelopathy group were better than those of patients in myeloradiculopathy group (P＜ 0.05).Conclusion Cervical artificial disc replacement has a satisfied long-term clinical efficacy with degenerative cervical canal stenosis.The efficacy of myelopathy and radiculopathy were better than myeloradiculopathy.

Objective:To introduce the application of a modified scrotoplasty for children with webbed penis.Methods:Retrospective study was carried out.The treatment results of 58 children with webbed penis in Department of Urology, the Affiliated Xuzhou Children′s Hospital of Xuzhou Medical University from June 2012 to April 2018 were analyzed.A total of 26 children with webbed penis and phimosis were treated with circumcision combined with modified scrotoplasty as modified scrotoplasty group, and 32 children with webbed penis and phimosis were treated with V-Y scrotoplasty as V-Y scrotoplasty group.The curative effect of webbed penis was compared between the two groups.Results:The operation time of V-Y scrotoplasty group and modified scrotoplasty group were (26.0±2.4) min and (28.0±3.2) min; the increased penis length in the horizontal position in the two groups were (0.30±0.06) cm and (0.40±0.06) cm; the score of parents′ satisfaction were (3.80±0.47) scores and (4.70±0.56) scores, there were significant differences in the average operation time, the increased penis length in the horizontal position and the score of parents′ satisfaction between the two groups ( t=2.703, 6.061, 6.652; all P<0.05). There was no significant difference in postoperative complications between the two groups ( P>0.05). Conclusions:Circumcision combined with modified scrotoplasty is used to repair webbed penis, which results in a good postoperative appearance, high parent satisfaction and easy clinical implementation.

Objective:To compare the effectiveness of modified Sugita and modified Shiraki in the treatment of severely concealed penis in children.Methods:A retrospective study was carried out on 72 children with congenital severely concealed penis, who were treated in the Department of Urology, Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University from September 2016 to June 2021.Among them, 32 cases were treated with modified Sugita (modified Sugita group) and 40 cases were treated with modified Shiraki (modified Shiraki group). There was no significant difference in the age, body mass and body mass index between the 2 groups (all P>0.05). The operation time, increased length of the exposed part of the penis after operation, score of parental satisfaction half a year after operation (obtained by using the Likert scale) and postoperative complications (classified according to the modified Clavien-Dindo classification) were compared between the 2 groups.Measurement data comparison between groups were performed by t test, counting data were analyzed by Chi- square test. Results:The operative time of the modified Sugita group and the modified Shiraki group were (50.00±8.03) min and (40.30±9.27) min, respectively.The operative time was significantly different between 2 groups ( t=4.107, P<0.05). The increased length of the exposed part of the penis after operation was (1.80±0.30) cm in the modified Sugita group and (1.90±0.33) cm in the modified Shiraki group, and no significant difference was found between the 2 groups ( P>0.05). Six months after operation, the parental satisfaction score was (4.60±0.56) points in the modified Sugita group and (4.60±0.59) points in the modified Shiraki group.There was no significant difference in the parental satisfaction score between 2 groups ( P>0.05). In 72 cases, only 5 cases had grade Ⅰ complications.Postoperative complications were also not significantly different between the 2 groups ( P>0.05). Conclusions:After treating severely concealed penis in children with modified Sugita and modified Shiraki, the penis has a good appearance.These two surgeries have high parental satisfaction and low complications.They are easy to implement in clinical practice.The operation time of modified Shiraki is shorter than that of modified Sugita.