Preparation and characterization of HBc virus like particles with site-directed coupling function / 生物工程学报

Hepatitis B virus core protein can self-assemble into icosahedral symmetrical viral-like particles (VLPs) in vitro, and display exogenous sequences repeatedly and densely on the surface. VLPs also have strong immunogenicity and biological activity. When the nanoparticles enter the body, they quickly induce specific humoral and cellular immune responses to exogenous antigens. In this study, we designed an HBc-VLPs that can be coupled with antigens at specific sites, and developed a set of efficient methods to prepare HBc-VLPs. Through site-specific mutation technology, the 80th amino acid of peptide was changed from Ala to Cys, a specific cross-linking site was inserted into the main immunodominant region of HBc-VLPs, and the prokaryotic expression vector pET28a(+)-hbc was constructed. After expression and purification, high purity HBc(A80C) monomer protein was assembled into HBc-VLPs nanoparticles in Phosphate Buffer. The results of particle size analysis show that the average particle size of nanoparticles was 29.8 nm. Transmission electron microscopy (TEM) showed that HBc-VLPs formed spherical particles with a particle size of about 30 nm, and its morphology was similar to that of natural HBV particles. The influenza virus antigen M2e peptide as model antigen was connected to Cys residue of HBc-VLPs by Sulfo-SMCC, an amino sulfhydryl bifunctional cross-linking agent, and M2e-HBc-VLPs model vaccine was prepared. The integrity of HBc-VLPs structure and the correct cross-linking of M2e were verified by cell fluorescence tracing. Animal immune experiments showed that the vaccine can effectively stimulate the production of antigen-specific IgG antibody in mice, which verified the effectiveness of the vaccine carrier HBc-VLPs. This study lays a foundation for the research of HBc-VLPs as vaccine vector, and help to promote the development of HBc-VLPs vaccine and the application of HBc-VLPs in other fields.

Application of indocyanine green in complex upper urinary tract repair surgery / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the clinical application of indocyanine green (ICG) visualization by near infrared fluorescence laparoscopy (NIFL) in complex upper urinary tract reconstructions surgery.@*METHODS@#This was a retrospective study of 7 patients who underwent complex surgeries of ureteral reconstruction between May 2019 and October 2019. There were 6 males and 1 female with the age ranging from 24 to 57 years (median age was 47 years). There were 5 cases of right ureteral strictures, of which 3 were proximal ureteral strictures and 2 were multiple and long ureteral strictures caused by radiotherapy. There were 2 cases of left ureteral strictures, of which 1 was ureteropelvic junction stricture and 1 was proximal ureteral stricture. There were 4 cases of secondary repair operations and 3 cases of primary operations. All the patients underwent laparoscopic surgery via the abdominal approach. ICG was injected into the ureter via nephrostomy tube during the operations, and the diseased ureter was identified by NIFL. Among the patients, 2 cases underwent IUPU (Institute of Urology, Peking University) modified ileal ureter replacement, 2 cases underwent ureteroureterostomy, 2 cases underwent appendiceal onlay flap ureteroplasty, and 1 case underwent lingual mucosa onlay flap ureteroplasty.@*RESULTS@#All the operations were successfully completed without open conversion. The localization and separation of ureteral lesions were completed under NIFL. The mean operative time was 187 (135-300) min. The duration of ureteral separation was 15-27 min, and the mean time was 18 min. The estimated blood loss was 15-200 mL, the mean estimated blood loss was 50 mL. There was one patient with ileal ureter replacement who had fever after surgery and responded well to antibiotics. The mean (range) length of postoperative hospital stay was 7 (6-10) days and no postoperative complications of a high grade (Clavien-Dindo Ⅲ and Ⅳ) occurred. Up to now, the mean follow-up duration was 9 (6-11) months, and no indocyanine green toxicity occurred. All D-J stents and nephrostomy were removed successfully 2 months after the operation. Ultrasound showed no obvious hydronephrosis, and CTU (computed tomography urography) showed that the urinary tract was unobstructed and the kidney function was normal.@*CONCLUSION@#The application of ICG in the complex upper urinary tract reconstructive surgery is a safe and easy method to help surgeon to identify the ureter which may reduce the risk of iatrogenic damage and protect the ureteral blood supply.

Research progress on Andersson lesion in ankylosing spondylitis / 中华外科杂志

Andersson lesion is a destructive vertebral or disco-vertebral lesion that occurs in the late stage of the ankylosing spondylitis. According to the etiology, these lesions are usually classified into localized lesions and extensive lesions. The history of ankylosing spondylitis and characteristic imaging is the key to the diagnosis of Andersson lesion. Conservative treatment may be effective for localized lesions. However, surgical intervention is often required for the failure of conservative treatment and extensive lesions. Currently, the optimal procedure for this problem is spinal osteotomy through pseudarthrosis and debridement via posterior-only approach.

Comparison of laparoscopic pyelolithotomy and percutaneous nephrolithotomy for renal pelvic stones larger than 2.5 cm / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the safety, efficacy and complications of laparoscopic pyelolithotomy (LPL) and percutaneous nephrolithotomy (PCNL) for treatment of renal pelvic stones larger than 2.5 cm.</p><p><b>METHODS</b>From 2011 to 2016, 32 patients underwent LPL and another 32 patients received PCNL for renal pelvic stones larger than 2.5 cm. The baseline characteristics of the patients, stone size, mean operative time, estimated blood loss, postoperative hospital stay, stone-free rate, postoperative analgesia, blood transfusion, and the intraoperative, early postoperative and long-term complications were compared between the two groups.</p><p><b>RESULTS</b>The baseline characteristics and stone size were comparable between the two groups. The mean operative time of LPL and PCNL was 117∓23.12 and 118.16∓25.45 min, respectively (P>0.05). The two groups showed significant differences in the mean estimated blood loss (63∓11.25 vs 122∓27.78 mL, P<0.01) and blood transfusion rate (0 vs 6.2%, P<0.01) but not in postoperative hospital stay (4.5∓1.34 vs 4.8∓2.2 days, P>0.05), stone-free rate (93.1% vs 87.5%, P>0.05) or the postoperative analgesia time (1.7∓0.5 and 1.9∓0.6 days, P>0.05). The incidence of intraoperative complications were significant lower in LPL group than in PCNL group (6.2% vs 25.0%, P<0.01), but the incidences of early postoperative complications (25.0% vs 34.4%, P>0.05) and long-term postoperative complications (9.4% vs 12.5%, P>0.05) were similar between them.</p><p><b>CONCLUSION</b>PCNL is the standard treatment for pelvic stones larger than 2.5 cm, but for urologists experienced with laparoscopic technique, LPL provides a feasible and safe option for management of such cases.</p>

Influence of BMP-7 on chondrocyte secretion and expression of Col-Ⅱ, AGG and Sox9 mRNA in porous tantalum-chondrocyte composites in vitro / 北京大学学报(医学版)

Objective:To study the influence of bone morphogenetic protein-7 ( BMP-7 ) on chondro-cyte secretion and expression of type Ⅱ collagen ( Col-Ⅱ) , aggrecan ( AGG ) and SRY-related high mobility group-box gene 9 ( Sox9 ) mRNA in porous tantalum-chondrocyte composites.Methods: The articular chondrocytes were isolated from 3-week-old New Zealand immature rabbits and identified.The 2nd generation of chondrocytes with 1 ×106/mL inoculate concentration was seeded in porous tantalum and divided into 4 groups, and control group ( tantalum/chondrocyte) , 50μg/L BMP-7 group (50μg/L BMP-7/tantalum/chondrocyte) , 100 μg/L BMP-7 group ( 100 μg/L BMP-7/tantalum/chondrocyte ) , and 200 μg/L BMP-7 group ( 200 μg/L BMP-7/tantalum/chondrocyte ) .The proliferation of chondro-cytes was measured by CCK-8 assay.The chondrocyte growth and morphology were observed by scanning electron microscopy ( SEM) .The synthesis of glycosaminoglycan ( GAG) in chondrocytes was tested by dimethyl methylene blue ( DMMB) colorimetric quantification method.Col-Ⅱ, AGG and Sox9 mRNA in chondrocytes were detected by real-time PCR.Results: The chondrocytes were spindle-shaped in 24 hours of primary cell culture and most cells became polygonal shaped in 4 days.The chondrocytes were affirmed by alcian blue, safranin O and Col-Ⅱimmunocytochemistry staining.The result of CCK-8 assay showed that the level of cell proliferation in 100 μg/L BMP-7 groups were higher than those in the other groups ( P<0 .05 ) .The chondrocytes implanted into porous tantalum scaffolds with BMP-7 had better functions, by which cytoplasmic processes developed and extended to the surface and inner of porous tan-talum by SEM observation.DMMB quantitative determination of GAG showed that GAG amount of chon-drocytes in 100 μg/L BMP-7 groups was significantly higher than those in the other groups ( P<0 .05 ) . The expressions of Col-Ⅱ, AGG and Sox9 mRNA in chondrocytes were up-regulated in the experimental groups, compared with the control group and the best effect appeared when concentration of BMP-7 was 200μg/L.(P<0.05).Conclusion:BMP-7/tantalum/chondrocytes composites enhanced in vitro chon-drocyte proliferation and extracellular matrix greatly, and can promote chondrogenic gene expression.

Effects of PPAR-gamma on the proliferation and glycolysis metabolism of prostate cancer cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the expression of the PPAR-gamma gene on the proliferation and glycolysis metabolism of prostate cancer cells.</p><p><b>METHODS</b>Using RNAi, we constructed lowly--expressed shRNA-PPARgamma adenoviruses and transfected them to PC3 prostate cancer cells, with blank vectors as controls. Then we detected the proliferation and apoptosis of the cells, glycolysis metabolism related genes and lactate accumulation by CCK-8 kit, and compared the results between the two groups.</p><p><b>RESULTS</b>Compared with the control group, the PPAR-gamma gene expression was obviously inhibited by RNAi in the PC3 cells, and its protein expression was reduced to (26.00 +/- 4.06)%. The proliferation inhibition rate was (39.5 +/- 4.92)% on the 2nd day, and the apoptosis rate was as high as (21.03 +/- 3.08)%. The glycolysis metabolism related gene products (Myc and Glut-1) were significantly decreased, and the lactate concentration was reduced to 69.71% of that of the controls on the 4th day. There were statistically significant differences in the above findings as compared with the control group (P < 0.01).</p><p><b>CONCLUSION</b>PPAR-gamma gene knockdown is expected to be a new way to treat prostate cancer.</p>

Preliminary study on biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma.</p><p><b>METHODS</b>Tumor samples were obtained from 5 patients, and then the human laryngeal carcinoma cells were cultured in vitro by primary tissue culture technique. Taking CD44 molecule as a marker to isolate CD(44)(+) subpopulation cells from laryngeal carcinoma cells for further study. CD(44)(+) and CD(44)(-) cells were cultured and observed fex their development. CD(44)(+) and CD(44)(-) cells were compared in their functional status (mRNA), cell cycles (G0/G1), their degree of differentiation (CK14 and Involucrin expression) and their morphologic character of the clone.</p><p><b>RESULTS</b>The percentages of CD(44)(+) cells were about 49.8% approximately 53.5% and the median was 51.3%. After culturing CD(44)(+) cells isolated from laryngeal carcinoma could proliferate and the percentage of CD(44)(+) remained the same. CD(44)(+) tumor cells contained much less RNA, more G0/G1 cells, expressed more CK14 protein and less Involucrin protein (less differentiated state). CD(44)(+) cells were multangular in shape with protuberances; CD(44)(-) cells showed a sharp and spindle feature. In comparison with CD(44)(-) cells, CD(44)(+) cells could create heterogeneous offspring by single cell culture of limiting dilution. By observing clone forming rate after single cell planting, it was found that the CD(44)(+) cells had stronger proliferation ability.</p><p><b>CONCLUSIONS</b>CD(44)(+) cells possess some characteristics of stem cells, laryngeal carcinoma stem cells maybe exist in CD(44)(+) cells.</p>

Detection of pim-1 mRNA in prostate cancer diagnosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pim-1 plays an important role in the apoptosis, proliferation, differentiation of cancer cells and progression of cancer. In this study we detected the expression of pim-1 mRNA in normal prostate, benign prostatic hyperplasia (BPH), and prostate cancer (PCa) and explored its diagnostic value for PCa.</p><p><b>METHODS</b>The prostate tissues were collected from 23 patients with PCa, 37 patients with BPH, and 3 healthy volunteers. Pim-1 mRNA expression levels in these samples were determined by the quantitative real-time PCR (QRT-PCR). The differences of expression were calculated based on a standard curve.</p><p><b>RESULTS</b>The ratio of pim-1 mRNA to beta-actin in the normal prostate, BPH, and PCa were 1.05 +/- 0.04, 2.57 +/- 0.74 and 4.45 +/-0.63, respectively. The differences among PCa, BPH and NT were significant (P < 0.05, respectively).</p><p><b>CONCLUSION</b>Detecting pim-1 mRNA expression by QRT-PCR provides a reliable metric for the diagnosis of PCa.</p>

cDNA macroarray for analysis of gene expression profiles in prostate cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Early diagnosis and timely treatment are important for improving therapeutic efficiency of prostate cancer. DNA array is a new bio-technology for disease diagnosis. This study was conducted to diagnose prostate cancer with cDNA macroarray and analysis gene expression profiles of some selective genes in prostate cancer.</p><p><b>METHODS</b>Total RNA was isolated from patients with prostate cancer and from normal people, and poly (A) RNA was further purified. Then it was analyzed for differentially expressed genes in prostate cancer and normal prostate by cDNA macroarray system.</p><p><b>RESULTS</b>There were different expressions in the nine prostate-associated specific genes in prostate cancer as compared with normal prostate, in which, 7 were significantly upregulated and 2 were down-regulated.</p><p><b>CONCLUSION</b>As a diagnostic approach at molecular level, the cDNA macroarray is an effectively diagnostic method for prostate cancer.</p>

Study on the strategy of Japanese encephalitis immunization using live attenuated vaccine combined with inactivated vaccine / 中华流行病学杂志

<p><b>OBJECTIVE</b>Using the advantages of Japanese encephalitis live attenuated and inactivated vaccine, to reduce the rate of immunization reaction and to increase the effect, we conducted a study on the strategy of immunization in Japanese encephalitis using live attenuated vaccine combined with inactivated vaccine.</p><p><b>METHODS</b>Observing the safety and immune effects of different groups.</p><p><b>RESULTS</b>Data on side effect showed that the rate of moderate and severe systematic reactions of the group who were inoculated with combined vaccine was 0.73%, with local reaction 1.46% while the combined rate of moderate and severe systematic reaction of the group who were inoculated with inactivated vaccine was 2.8%. Under the detection of serum neutralizing antibody, the GMT rose from 1:1.05 - 1:3.35 before vaccination to 1:47.34 - 1:101.30 after vaccination in the different groups. Neutralizing antibody was detected in 97.67% of the combined group. There was a significant difference by comparing neutralizing antibody seroconversion rate of the combined group with the inactivated group (chi(2) = 3.89, P < 0.05), but no significant difference with attenuated group (chi(2) = 0.74, P > 0.05).</p><p><b>CONCLUSION</b>Results showed that in children who previously had been immunized with two doses of inactivated vaccine, the booster administration of live attenuated vaccine was both effective and safe.</p>

The evaluation of angiographic features of intracranial arteries in patients with ischemic cerebral stroke / 介入放射学杂志

Ischemic cerebral stroke is one of the diseases posing great threat to human health.It is of great value to have a correct evaluation and interpretation of cerebral angiography in interventional treatment of ischemic stroke together with efficacy.The authors reviewed the direct signs and collateral circulation of ischemic cerebral stroke and the evaluation of cerebral angiography in recent literature.