Dissection of Cellular Communication between Human Primary Osteoblasts and Bone Marrow Mesenchymal Stem Cells in Osteoarthritis at Single-Cell Resolution

Background and Objectives@#Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and playimportant role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells. @*Methods@#and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identifynew cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs. @*Conclusions@#Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.

Effects of Dasatinib on the Maturation of Monocyte-Derived Dendritic Cells Derived from Healthy Donors and Chronic Myelogenous Leukemia Patients / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of dasatinib on the maturation of monocyte-derived dendritic cells (moDCs) derived from healthy donors (HDs) and chronic myelogenous leukemia (CML) patients.@*METHODS@#Peripheral blood mononuclear cells (PBMCs) were isolated from HDs (n=10) and CML patients (n=10) who had got the remission of MR4.5 with imatinib treatment. The generation of moDCs from PBMCs was completed after 7 days of incubation in DC I culture medium, and another 3 days of incubation in DC II culture medium with or without 25 nmol/L dasatinib. On the 10th day, cells were harvested and expression of molecules of maturation related marker were assessed by flow cytometry. The CD80+CD86+ cell population in total cells was gated as DCs in the fluorescence-activated cell storting (FACS) analyzing system, then the expression of CD83, CD40 or HLA-DR in this population was analyzed respectively.@*RESULTS@#The proportion of CD80+CD86+ cells in total cells didn't show a statistical difference between HD group and patient group (89.46%±9.70% vs 87.39%±9.34%, P=0.690). Dasatinib significantly enhanced the expression of the surface marker CD40 (P=0.008) and HLA-DR (P=0.028) on moDCs derived from HDs compared with the control group, while the expression of CD83 on moDCs didn't show a significant difference between dasatinib group and the control group (P=0.428). Meanwhile, dasatinib significantly enhanced the expression of the surface marker CD40 (P=0.023), CD83 (P=0.038) and HLA-DR (P=0.001) on moDCs derived from patients compared with the control group.@*CONCLUSION@#For CML patients, the same high proportion of moDCs as HDs can be induced in vitro, which provides a basis for the application of DC-based immunotherapy strategy. Dasatinib at the concentration of 25 nmol/L can efficiently promote the maturation of moDCs derived from HDs and CML patients in vitro. Dasatinib shows potential as a DC adjuvant to be applied in DC-based immunotherapy strategies, such as DC vaccine and DC cell-therapy.

Management of anastomotic leakage after laparoscopic surgery for adenocarcinoma of esophagogastric junction / 中华胃肠外科杂志

The incidence of adenocarcinoma of esophagogastric junction (AEG) is increasing at home and abroad. Laparoscopic surgery has gradually become the main means of surgical treatment of this kind of tumor. However, due to the special anatomical position of the tumor, the high position away from the broken esophagus and the narrow space in the mediastinum, laparoscopic anastomosis has the characteristics of difficult anastomosis and high anastomosis position. There is a high risk of anastomotic leakage after operation, which may cause serious consequences. Early identification of anastomotic leakage and unobstructed drainage by various means are the key to treatment. With the development of endoscopic technology, endoscopic methods such as covered stent and vacuum-assisted closure further improve the treatment efficacy. As a salvage measure, surgical treatment can achieve good treatment outcome, while accompanied by risk of complications and mortality, so we must strictly grasp the indications.

Functional evaluation after function-preserving gastrectomy / 中华胃肠外科杂志

With the increase of people's health awareness and the progress of medical diagostic technology in recent years, the diagnosis rate of early gastric cancer is increasing year by year. Although radical surgery has good efficacy, how to maximize the preservation of the normal anatomy and function of the stomach and improve the quality of life of patients in the pursuit of radical surgery has become a more important issue in the treatment of early gastric cancer. Under the condition of ensuring radical lymph node dissection, function-preserving gastrectomy can fully preserve gastric function by reducing the resection extent and preserving the pylorus and the vagus nerve, which has advantage of improving quality of life and has great potential in the treatment of early gastric cancer. However, there is no functional evaluation standard for function-preserving gastrectomy at present. Most of the patients are evaluated by quality of life scale, which is relatively subjective. Even though the evaluation of endoscopy, hematology and other objective means can indicate the benefit degree in quality of life brought by functional reconstruction, the evidence level is limited. Therefore, this paper discusses the research status of function-preserving gastrectomy evaluation, postoperative complications, postoperative nutritional status, auxiliary examination and other items in the evaluation of gastric function, and analyzes the prospects of research direction in this field.

Chemical Constituents in Endophytic Fungus Nigrospora oryzae from Cordyceps / 中国实验方剂学杂志

Objective: To study the chemical constituents in theendophytic fungus Nigrospora oryzae from Cordyceps. Method: It was cultured with brownrice,and isolated and purified by chromatographic procedures, and the compounds were identified by NMR,ESI-MS and other spectral methods. Result: Totally 15 compounds were identified as mellein (1),linoleic acid (2),2-(2-hydroxyethyl)phenol (3),(3R)-mellein methyl ether (4),(3R,4S)-4-hydroxymellein (5),2-hydroxybenzaldehyde (6),3-phenylpropane-1,2-diol (7),1-phenyl-1,2-ethanediol (8),cyclo-(R-Prommmm-S-Ile) (9),cyclo-(D-Pro-L-Leu) (10),2-hydroxybenzaldehyde (11),4-hydroxy-8-O-methylmellein (12),cyclo-(S-Pro-S-Phe) (13),cyclo-(D-Pro-L-Ile) (14),cyclo-(D-Pro-L-Leu) (15). Conclusion: Compounds 1-15 were isolated from this fungus for the first time.

Cytoprotective effect of p62/Nrf2 signaling pathway / 药学学报

The Nrf2-Keap1-ARE pathway is an important signaling axis that functions to protect cells against oxidative stress and harmful chemicals through the induction of cytoprotective genes. The maintenance and protective role of Nrf2 pathway has been recognized as a means for chemoprevention. On the other hand, constitutive activation of Nrf2, due to somatic mutations of genes that control Nrf2 degradation, promotes carcinogenesis and imparts chemoresistance to cancer cells. Autophagy is another tightly regulated complex cellular process that functions as a cellular quality control system to remove damaged proteins or organelles. Recently, these two cellular pathways were shown to intersect through the direct interaction between p62 (an autophagy adaptor protein) and Keap1. Dysregulation of autophagy was shown to result in prolonged activation of Nrf2 in a p62-dependent manner, which is associated with the pathogenesis and therapies of several human diseases including cancer. In this review, we discuss the molecular mechanisms of p62-mediated Nrf2 signaling pathway, with a special emphasis on their impact on nervous system disease, cardiovascular disease and cancer.

Clinical outcomes of using nonmodular tapered long stem in femoral revision with Paprosky Ⅲ bone defect / 医学研究生学报

Objective It is a great challenge to perform hip revision surgery especially with severe proximal femoral bone de -fect.This study aims to discuss the short-medium clinical outcomes of using nonmodular full-coated tapered stem in femoral revision with Paprosky Ⅲ bone defect. Methods From May 2011 to October 2015, 58 patients(58 hips)diagnosed as ostiolysis/aseptic loosening with severe femoral bone defect(Paprosky Ⅲ)were treated with hip revision surgery.The cases of septic loosening and tumor were excluded.Harris scoring system and pain visual analogue scale(VAS)were used pre-and post-operatively to evaluate the function of hip joints and the pain level.Radiograph was used to evaluate loosening,subsidence and bone healing of the prosthesis. Results All patients were followed-up with 24-67 months,the mean follow up time was 49.3±12.7 months.None of the patients showed infec-tion or surrounding bone fracture.Dislocation happened in one case after surgery and was treated with manipulative reduction without re -currence.One case had progressive subsidence which led to aseptic loosening and was treated with re-revision.By the end of last follow-up,Haris Score was improved from preoperative 41.6±6.9 points to 83.1±5.7 points, VAS score decreased from preoperative 6.7 ±1.6 points to 2.5 ±1.2 points,(P<0.05). Conclusion Nonmodular full-coated tapered stem can obtain good clinical effect in femoral revision with severe proximal bone defect.

Association of CT perfusion imaging with plasma levels of TGF-β1 and VEGF in patients with NSCLC

OBJECTIVE@#To study the association of CT perfusion imaging parameters with plasma level of transforming growth factor-β1 (TGF-β1) and vascular endothelial growth (VEGF) in patients with non small cell lung cancer (NSCLC).@*METHODS@#A total of 67 patients with NSCLC (NSCLC group) and 64 patients with benign lesion (control group) were given with CT perfusion imaging to obtain blood flow, blood volume, mean transit time, time to peal and permeability surface through CT perfusion software. The plasma levels of TGF-β1 and VEGF were tested by ELISA. The relationship between plasma levels of TGF-β1, VEGF and CT perfusion imaging parameters were analyzed.@*RESULTS@#CT perfusion imaging parameters and the plasma levels of TGF-β1 and VEGF of NSCLC group were significantly higher than the control group (P < 0.05), while CT perfusion parameters and the levels of TGF-β1 and VEGF in NSCLC group showed significant difference in different tumor node metastasis stages (P < 0.05). Correlation analysis showed that the level of plasma TGF-β1 and VEGF were positively correlated with blood flow, blood volume, and mean transit time (P < 0.05), and negatively correlated with time to peal (P < 0.05). There was no significant correlation between TGF-β1 and VEGF with the permeability surface.@*CONCLUSIONS@#CT perfusion imaging parameters in patients with NSCLC is closely associated with plasma TGF-β1, VEGF and its biological characteristics. CT perfusion imaging is a convenient method to detect tumor blood perfusion.

Association of CT perfusion imaging with plasma levels of TGF-β1 and VEGF in patients with NSCLC

Objective: To study the association of CT perfusion imaging parameters with plasma level of transforming growth factor-β1 (TGF-β1) and vascular endothelial growth (VEGF) in patients with non small cell lung cancer (NSCLC). Methods: A total of 67 patients with NSCLC (NSCLC group) and 64 patients with benign lesion (control group) were given with CT perfusion imaging to obtain blood flow, blood volume, mean transit time, time to peal and permeability surface through CT perfusion software. The plasma levels of TGF-β1 and VEGF were tested by ELISA. The relationship between plasma levels of TGF-β1, VEGF and CT perfusion imaging parameters were analyzed. Results: CT perfusion imaging parameters and the plasma levels of TGF-β1 and VEGF of NSCLC group were significantly higher than the control group (P < 0.05), while CT perfusion parameters and the levels of TGF-β1 and VEGF in NSCLC group showed significant difference in different tumor node metastasis stages (P < 0.05). Correlation analysis showed that the level of plasma TGF-β1 and VEGF were positively correlated with blood flow, blood volume, and mean transit time (P < 0.05), and negatively correlated with time to peal (P < 0.05). There was no significant correlation between TGF-β1 and VEGF with the permeability surface. Conclusions: CT perfusion imaging parameters in patients with NSCLC is closely associated with plasma TGF-β1, VEGF and its biological characteristics. CT perfusion imaging is a convenient method to detect tumor blood perfusion.

Study on suitable harvest time of Dendrobium officinale in Yunnan province / 中国中药杂志

In order to determine the suitable harvest time of Dendrobium officinale from different regions in Yunnan province, the drying rate, mannose and glucose peak area ratio, extract, contents of polysaccharide and mannose of D. officinale samples collected from six producing areas in Ynnnan province were determined. The results indicate that drying rate and the contents of polysaccharide and mannose arrived the peak from January to April, extract reached a higher content from September to December, and mannose and glucose peak area ratio from October to February of the coming met the requirment of the Chinese Pharmacopoeia. Hence, the suitable harvesting time of D. officinale in Yunnan province is from December to February of the coming year,according to the experimental results and the request of the Chinese Pharmacopoeia.

Neonatal hyperthyroidism: a case report and literature review / 南方医科大学学报

We report a case of neonatal thyrotoxicosis with concurrent respiratory failure in an infant born to a mother with Graves' disease and review the published literature describing neonatal hyperthyroidism. The male infant who was born by spontaneous delivery at 35 weeks of gestational age presented with fever, tachycardia and tachypnea at rest on day 11 after birth, and developed severe apnea on day 14. Thyroid function studies revealed hyperthyroidism in the infant, and his mother was confirmed to have Grave's disease during pregnancy. Literature review showed that among the 33 infants with similar conditions, tachycardia, tachypnea and poor weight gain were the most distinct clinical features of congenital hyperthyroidism. Accurate diagnosis of Graves' disease in the mother during pregnancy and awareness of the clinical presentations of neonatal hyperthyroidism are key to reducing missed diagnosis or misdiagnosis of neonatal hyperthyroidism.

Application of 3D-computed tomography angiography technology in large meningioma resection

OBJECTIVE@#To discuss the role of 3D-computed tomography angiography (3D-CTA) technology in reducing injuries of large meningioma surgery.@*METHODS@#3D-CTA preoperative examinations were done in 473 patients with large meningioma (simulated group). The images were analyzed by 3D post-processing workstation. By observing the major intracranial blood vessels, venous sinus, and the compression and invasion pattern in the nerve region, assessing risk level of the surgery, simulating the surgical procedures, the surgical removal plan, surgical routes and tumor blood-supplying artery embolisation plan were performed. Two hundred and fifty seven large meningioma patients who didn't underwent 3D-CTA preoperative examination served as control group. The incidence of postoperative complications, intraoperative blood transfusion and the operation time were compared between these two groups.@*RESULTS@#Compared with the control group, the Simpson's grade I and II resection rate was 80.3% (380/473), similar with that of the control (81.3%, 209/257). The incidence of postoperative complications in 3D-CTA simulated group was 37.0% which was significantly lower than that (48.2%) of the control (P<0.01). The intraoperative blood supply for simulated group and the control was (523.4±208.1) mL and (592.0±263.3) mL, respectively, with significant difference between two groups (P<0.01). And the operation time [(314.8±106.3)] min was significantly lower in simulated group than that in the control [(358.4±147.9) min] (P<0.01).@*CONCLUSION@#Application of 3D-CTA imaging technology in risk level assessment before large-scaled meningioma resection could assist in the rational planning of tumor resectin, surgical routes, and is helpful in reducing injuries and complications and enhancing the prognosis of the patients.

Study of the correlation between polycyclic aromatic hydrocarbons exposure during pregnancy and neonatal neurobehavioral development in Taiyuan and Changzhi cities / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To compare the difference of polycyclic aromatic hydrocarbons (PAHs) levels in the urban air and the scores of Neonatal Behavioral Neurological Assessment (NBNA) between Taiyuan and Changzhi cities and to explore the effects of PAHs in the urban air during pregnancy on neonatal behavioral neurological development.</p><p><b>METHODS</b>High-performance liquid chromatography (HPLC) with subsequent fluorescence detection was used to determine the PAHs levels in the cooperational hospitals in Changzhi and Taiyuan cities and the urinary 1-hydroxypyrene levels of the 297 pregnant women living Changzhi and Taiyuan cities during Nov. 2009 to May 2010. NBNA was used to determine the development of neonatal neural behavior. The differences of PAHs levels in the urban air, the pregnant women urinary 1-hydroxypyrene levels and NBNA scores between Taiyuan and Changzhi were compared.</p><p><b>RESULTS</b>There are significant differences of levels of pyrene, benz [a] anthracene, Chrysene, benz [a] pyrene, dibenz [a, h] anthracene in the urban air between Taiyuan and Changzhi (P < 0.10). The median of urinary 1-hydroxypyrene levels in pregnant women of Taiyuan was 1.140 microg/mmolCr, (P25 was 0.457 microg/mmolCr, P75 was 2.678 microg/mmolCr), the median of urinary 1-hydroxypyrene levels in pregnant women of Changzhi was 0.761 microg/mmolCr, (P25 was 0.133 microg/mmolCr, P75 was 2.095 microg/mmolCr). There are significant differences of urinary 1-hydroxypyrene levels in pregnant women between Taiyuan and Changzhi (t = -3.140, P = 0.002). There are significant differences of the NBNA scores, capacity scores, passive muscle tension scores, active muscle tension scores and general assessment scores between Taiyuan and Changzhi (P < 0.10). There was correlation between NBNA scores and urinary 1-hydroxypyrene level in pregnant women.</p><p><b>CONCLUSION</b>The PAHs in the urban air during pregnancy may adversely affect the neonatal neurobehavioral development.</p>

Prenatal screening and diagnosis of genetic deafness by microarray / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate a microarray-based mutation screening method for genetic deafness and its application in prenatal diagnosis.</p><p><b>METHODS</b>Mutation screening of common deafness genes was performed in pregnant women and volunteers spouses. Nine common mutations in four major deafness genes, GJB2, GJB3, SLC26A4 and mitochondrial 12S rRNA, were detected simultaneously by a microarray-based method. Genetic counseling was given based on their testing results.</p><p><b>RESULTS</b>5.11% of pregnant women carried at least one mutation. Among them, seven carried mutation in the mitochondria 12S rRNA gene and were offered aminoglycoside-induced ototoxicity warning. For other mutation carriers of GJB2 or SLC26A4 genes, additional mutation screening was performed in their husbands by direct sequencing. A total of 20 couples were at risk of giving birth to children with genetic deafness. Of five couples who selected to undergo prenatal diagnostic testing of the fetus, four were diagnosed as wild type or heterozygous for the tested genes and one as p.V37I/c.235delC compound heterozygous for GJB2.</p><p><b>CONCLUSIONS</b>DNA microarray is a quick, easy and reliable method to screen mutations in genetic deafness genes. Application of this method in prenatal screening and diagnosis might effectively reduce the occurrence of genetic deafness.</p>

Association of retinal diseases with epigenetics mechanism / 中华实验眼科杂志

Epigenetics refers to the changes of heritable gene expression without modification of DNA sequences which is one of major research interest of human genome study post gene era. Epigenetics may play an important role in the pathogenesis of retinal diseases. Epigenetics modification in the regulation of gene expression and function mainly includes DNA methylation,histone acetylation and non-coding RNA such as miRNA. The implication of epigenetics has been suggested with the pathogenesis of retinal fibrosis, retinitis pigmentosa, age-related macular degeneration and diabetic retinopathy. In addition, the epigenetics drug has potential to be used in the treatment of retinal diseases.

Synopsis of epigenetics / 中华实验眼科杂志

Epigenetics has become a hot topic in biomedical research in last a few years.Epigenetics refers to as epigenome,and it is interpreted as epigenetic changes (DNA methylation,histone modifications and non-coding RNAs such as microRNAs) in the regulation of genes expression on chromatin-based pathways rather than a single gene.Epigenetic factors such as DNA methylation,histone modification and miRNA serve as an interface for environmental stimuli and interacts each other to regulate gene expression and control phenotypes of cell.All of those epigenetic factors are essential in the keeping homeostasis of human body and contribute to many biologic conditions.In addition,the application of epigenetic drugs has been come into clinical trials especially in the treatment of cancers,therefore the understanding the mechanism of epigenetics in the development of human diseases and the therapeutic potential of epigenetic regulators in the treatment of human disease will bring us into the epigenetics era in biomedical research.

Effect of proteasome inhibitor MG-132 on L1210 cell apoptosis and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the effect of proteasome inhibitor MG-132 on apoptosis of L1210 cells and its mechanism. L1210 cells were treated with MG-132 of different concentrations (0, 2.5, 5, 10, 10 micromol/L). Cell viability was tested by MTT assay, apoptosis rate was detected by using flow cytometry, activity of caspase 3 was detected by colorimetry, the expression of NF-kappaB nuclear protein was detected by Western blot. The results showed that the growth inhibition of L1210 cells treated for a same time (24 hours) was enhanced along with increasing of MG-132 concentrations (0, 2.5, 5, 10, 20 micromol/L); the inhibitory rate, apoptosis rate and activity of caspase 3 increased also along with raising of MG-132 concentrations; while the expression of NF-kappaB nuclear protein decreased along with raising of GM-132 concentrations. It is concluded that MG-132 can induce the apoptosis of L1210. The mechanism of apoptosis may be related to the down-regulation of the expression of NF-kappaB and the activation of caspase 3.

Influence of suppression of Epstein-Barr Virus-encoded latent membrane protein 1 by rAAV vector mediated RNA interference on metastatic ability of nasopharyngeal cancer cells in vivo / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP-1) on the tumor growth and metastasis of nasopharyngeal carcinoma (NPC) cells in vivo and its possible mechanism.</p><p><b>METHODS</b>To construct two recombinant adeno-associated virus (rAAV): rAAV-shRNA-LMP-1 and rAAV-EGFP (enhanced green fluorescent protein). Multiplicity of infection (MOI) was confirmed by using different titre of rAAV-EGFP to transfect a NPC cell line, C666-1. Then C666-1 cells were transfected by rAAV-shRNA-LMP-1 at MOI titre and the inhibiting efficiency of target gene's expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR). C666-1 cells treated by RNAi on LMP-1 in vitro was directly inoculated into the liver via laparotomy under direct vision to establish the animal model of NPC xenograft in liver and lung metastasis from the liver. The biological effect and its mechanism after "gene silencing" of LMP-1 on NPC cells tumorigenesis and metastasis were observed by the primary intrahepatic tumor formation and lung metastasis and the expression of matrix metalloproteinases-9 (MMP-9) revealed by immunohistochemistry.</p><p><b>RESULTS</b>The transfection efficiency was higher than 95% with 5 x 10(4) virus genome (v.g)/cell with rAAV-EGFP. The expression of target gene was inhibited more than 90%, assessed by RT-PCR after transfection with rAAV-shRNA-LMP-1 at a dose of 5 x 10(4) v.g/cell. The primary tumor volume implanted in the liver of rAAV-shRNA-LMP-1 treatment was (0.2527 +/- 0.1152) cm3, with no significant difference in comparrison with rAAV-EGFP control group [(0.2533 +/- 0.0754) cm3, P>0.05]. But the rate of intrahepatic tumor formation was 50.0% and the rate of lung metastasis was 33.3% of the rAAV-shRNA-LMP-1 group, significantly lower than those in the rAAV-EGFP group (P<0.05). The survival time (15.50 +/- 2.47) d of rAAV-shRNA-LMP-1 group was significantly longer than that in the rAAV-EGFP group [(11.50 +/- 1.22) d, P<0.05]. Immunohistochemistry indicated suppression of LMP-1 by rAAV-shRNA-LMP-1 can down-regulate the expression of MMP-9 significantly.</p><p><b>CONCLUSION</b>The expression of LMP-1 can be suppressed effectively by rAAV mediated RNA interference. The suppression of LMP-1 expression has no effect on cell growth but can inhibit the metastasis in vivo, probably through down-regulating the expression of MMP-9.</p>

Effect of rAAV-mediated RNA interference against EB virus-encoded latent membrane protein 1 on nasopharyngeal carcinoma cell proliferation and metastasis in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the role of Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP-1) in nasopharyngeal carcinoma (NPC) cell growth and metastasis in vivo by recombinant adeno-associated virus (rAAV)-mediated RNA interference (RNAi).</p><p><b>METHODS</b>Specific small hairpin RNA (shRNA) targeting EBV-LMP-1 gene was designed and synthesized to construct two rAAV vectors rAAV-shRNA-LMP-1 and rAAV-EGFP. The multiplicity of infection (MOI) was confirmed using different titers of rAAV-EGFP to transfect the NPC cell line C666-1. The C666-1 cells were transfected by rAAV-shRNA-LMP-1 at the optimal MOI titer and the inhibition efficiency of the target gene expression was determined with RT-PCR. The C666-1 cells with RNAi of LMP-1 gene were injected into nude mouse liver via laparotomy to establish the animal model of hepatic and lung metastases of NPC cells. The metastases of the C666-1 cells in the liver and lungs were observed to assess the effect of LMP-1 gene silencing on the tumorigenic and metastatic potentials of the cells in vivo.</p><p><b>RESULTS</b>The transfection efficiency of 5 x 10(4) virus genome/cell rAAV-EGFP exceeded 95%. The expression of the target gene was suppressed by over 90% as shown by RT-PCR after transfection with rAAV-shRNA-LMP-1 at 5 x 10(4) virus genome/cell. Animal experiments showed that compared with rAAV-EGFP, rAAV-shRNA-LMP-1 transfection did not reduce the primary tumor volume implanted into the liver, but significantly inhibited the intrahepatic and lung metastases of the NPC cells.</p><p><b>CONCLUSION</b>LMP-1 expression can be suppressed effectively by rAAV-mediated RNAi, and LMP-1 suppression does not obviously affect the tumor cell growth but can inhibit their metastasis in vivo.</p>

The use of uncemented extensively porous-coated femoral components in the management of Vancouver type B2 periprosthetic femoral fractures / 中华外科杂志

<p><b>OBJECTIVE</b>To assess the outcome of patients with Vancouver type B2 periprosthetic fractures treated with femoral revision using an uncemented extensively porous-coated implant.</p><p><b>METHODS</b>From October 2002 to May 2007, a retrospective clinical and radiographic assessment of 10 patients with Vancouver type B2 periprosthetic fractures treated with femoral revision was evaluated, among which 4 was cement fixation, 6 was non-cement fixation in the index procedure.</p><p><b>RESULTS</b>The mean follow-up for these patients was 44 months (range, 12 to 67 months). The mean Harris hip score was 87.6 points. All 10 reconstructions showed solid fracture union and a stable prosthesis, 8 was bone ingrowth, 2 was fibrous ingrowth. Complications in 4 patients included thigh pain in 1, stress shield in 3.</p><p><b>CONCLUSIONS</b>Vancouver type B2 periprosthetic fractures treated with uncemented extensively porous-coated femoral stems incorporate distally allowing stable fixation.</p>