Fluorescence in situ hybridization based on a panel of probes for detection of common cytogenetic abnormalities in multiple myeloma / 实用医学杂志

Objective To evaluate the advantages of plasma cell enrichment combined with fluorescence in situ hybridization (FISH) based on a panel of probes by the conventional cytogenetic (CC) analysis.Methods Fresh heparinized bone marrow samples were collected by bone marrow biopsy.Plasma cells were enriched in BM samples using a magnetic cell-sorting procedure to select CD138+ cells.The common chromosome abnormalities of MM were detected by FISH based on a panel of probes and CC analysis after short-term culture of the BM cells,in order to compare the differences between these two methods for the frequency of common cytogenetic abnormalities.Results 72 of 95 (75.8％) MM patients were found to carry clonal chromosome abnormalities by FISH.And RB 1 deletion was the highest at 44.2％ (42/95) followed by CKS1B (1q21) amplification (42.1％).The frequencies of CDKN2C (1p32) deletion,TP53 deletion,IGH/CCND1 and IGH/FGFR3 were 8.4％ (8/95),12.6％ (12/95),14.7％ (14/ 95) and 14.7％ (14/95),respectively.IGH/MAF was negative.Thirty-two of 95 (33.7％) patients were found to carry clonal aberrations by CC analysis.The frequency of chromosome abnormalities detected by FISH was significantly higher than CC analysis (75.8％ vs 33.7％,P =0.000).Conclusion Plasma cell enrichment combined with FISH based on a panel of probes can greatly increase the frequency of chromosome abnormalities,which provides cytogenetic basis for risk stratification and prognosis of MM patients.

Efficacy comparison between targeted and conventional percutaneous vertebroplasty of osteoporotic vertebral compression fractures / 中华创伤杂志

Objective To compare the clinical outcomes between conventional percutaneous vertebroplasty (PVP) and targeted PVP in the treatment of osteoporotic vertebral compression fractures (OVCFs).Methods A retrospective cohort study was designed to review 215 cases of single level OVCFs hospitalized between January 2014 and December 2015.According to the procedure techniques,the patients were assigned to targeted PVP group (89 cases) and conventional PVP group (126 cases) which was further divided into sufficient filled subgroup (110 cases) and insufficient filled subgroup (16 cases) on basis of cement distribution.Key techniques of targeted PVP included accurate needle insertion to fractured area and cement injection using a push rob with a side opening.Operating time,cement injection volume,rate and types of cement leakage,cement distribution in the fractured area and visual analogue score (VAS) of back pain were compared between the two groups.Results Operating time in targeted PVP group was longer than that in conventional PVP group (P ＜ 0.05).There were no significant differences in cement injection volume and rate and types of cement leakage between the two groups (P ＞ 0.05).None in targeted PVP group showed insufficient cement distribution in fractured area,while 16 cases (12.7％) in conventional PVP group (P ＜ 0.05).No significant differences in preoperative VAS of back pain existed among targeted PVP group,sufficient subgroup and insufficient subgroup (P ＞ 0.05).VAS of back pain was significantly decreased after PVP in three groups (P ＜ 0.05).Difference in postoperative VAS of back pain between targeted PVP group and sufficient filled subgroup was insignificant (P ＞0.05).However,postoperative VAS of back pain in insufficient filled subgroup was significantly increased compared with targeted PVP group and sufficient filled subgroup (P ＜ 0.05).Conclusion Targeted PVP provides sufficient cement to fill the fractured area and decreases incidence of unsatisfactory clinical outcome compared with traditional PVP,indicating a secure and effective new technique in the treatment of OVCFs.

Research advancement of calcium phosphate and calcium sulfate scaffolds in bone tissue engineering / 中国组织工程研究

BACKGROUND: It is a hotspot that calcium phosphate and calcium sulphate as the main ingredients are combined with one or more other materials to improve or increase the performance of bone tissue engineering scaffolds. OBJECTIVE: To introduce the research advance of these two kinds of scaffolds in bone tissue engineering. METHODS: The articles related to the bone tissue engineering published during January 2000 to June 2015 were retrieved from CNKI and PubMed databases by computer. The key words were “bone tissue engineering, scaffold, calcium phosphate, calcium sulphate, vascularization” in Chinese and English, respectively. ESULTS AND CONCLUSION: Calcium phosphate and calcium sulfate are characterized as having good biocompatibility, biodegradability, osteoconductivity and complete bone substitutability. However, single use of calcium phosphate or calcium sulfate scaffold has certain disadvantages, both of which are difficult to ful y meet the requirements of the bone defect repair. Improvement can be acquired in the mechanical strength, injectability and biodegradability, as wel as drug-loading and pro-angiogenesis of the scaffold in combination with other materials. In the basal and clinical research, we should explore and develop ideal scaffolds in on the basis of therapeutic aim. However, most of the scaffold studies are stil at the extracorporeal and animal experiment stage, and the comparative studies on composite scaffolds and optimal proportion of those composite scaffolds stil need to be further investigated.

Stability and interbody fusion of augmented pedicle screws with bone cement for lumbar spondylolisthesis accompanied with osteoporosis / 中国组织工程研究

BACKGROUND: In lumbar spondylolisthesis patients with severe osteoporosis, screw is easily loose and pul s out during reposition, or loss of reduction and internal fixation failure easily occur after repair. Therefore, it is very important to elevate the intensity of pedicle screw fixation during repair. At present, few studies concern application of bone cement screw enhancement technology in lumbar spondylolisthesis patients with osteoporosis. OBJECTIVE: To investigate the clinical value of augmented pedicle screw with polymethylmethacrylate for lumbar spondylolisthesis accompanied with osteoporosis. METHODS: From June 2009 to June 2011, 27 patients suffering from lumbar spondylolisthesis accompanied with osteoporosis were included in this retrospective study. These patients received augmented pedicle screw with polymethylmethacrylate. The levels of disability and pain were evaluated by Oswestry Disability Index and visual analog scale. The internal fixation and fusion were evaluated by radiological findings. Al complications were recorded. RESULTS AND CONCLUSION: Al cases were fol owed up for 15-37 months. Oswestry Disability Index and visual analog scale scores were significantly better in final fol ow-up than that pre-treatment (P < 0.05). Imaging results revealed that bone cement tightly connected to bone interface. The position of screw and bone cement was good. Symptomatic bone cement leakage was not found. No fixation failure was detected during final fol ow-up. Al patients achieved interbody fusion. These results suggested that polymethylmethacrylate bone cement could increase the gripping force of the pedicle screw in osteoporotic vertebral body. It is safe and effective to treat spondylolisthesis accompanied with osteoporosis with augmented pedicle screws. Satisfactory fixation stability and interbody fusion can be obtained.

Gelatin sponge containing prednisolone acetate relieves radicular pain following posterior lumbar interbody fusion / 中国组织工程研究

BACKGROUND:Posterior lumbar interbody fusion can thoroughly decompress the central canal, which is the common surgical technique for the central type of lumbar disc herniation with intervertebral instability at low lumbar segment. However, due to the regular traction on dural sac and nerve root in the operation, lower limb radicular pain in the early stage is inevitable. OBJECTIVE:To evaluate the effect of local use of gelatin sponge containing prednisolone acetate around the nerve roots after posterior lumbar interbody fusion on lower limb radicular pain. METHED:Sixty-three cases of lumbar disc herniation with degenerative instability were devided into treatment group (n=21) and control group (n=42) based on the type of implants. Gelatin sponge containing prednisolone acetate was implanted into patients in the treatment group after posterior lumbar interbody fusion, while pure gelatin sponge was implanted into patients in the control group. RESULTS AND CONCLUSION:Compared with the control group, radicular pain in the treatment group was significantly relieved within 1 week after surgery. The visual analog scale score and Oswestry disability index score were similar between the two groups. There were three cases of radicular pain recurrence in the control group, but no incision infection and epidural hematoma after surgery in both two groups. In conclusion, local use of gelatin sponge containing prednisolone acetate around the nerve roots can significantly relieve lower limb radicular pain in the early stage after posterior lumbar interbody fusion in lumbar disc herniation, contributing to early rehabilitation exercise and patient satisfaction outcomes.

Discussion on converging the doctor workstation with the epidemic information monitoring and reporting system / 中华医院管理杂志

The networked epidemic information reporting management system and reporting mechanism of the hospital was called into play,embedding the reporting system into the outpatient and resident workstations of the hospital's IT management system.Such a networked platform enables the overall coverage and real-time reporting,monitoring and prewarning of hospital-wide epidemic information,providing reliable support for control and prevention of hospital epidemic control.

Clinical studies of risperidone in the treatment of the mental symptoms after frontotemporal brain contu-sion / 中国神经精神疾病杂志

Objective To examine the effect of risperidone on the mental symptoms after frontotemporal brain contu?sion. Methods Sixty cases with mental symptoms after frontotemporal brain contusion were recruited from Jan 2009 to Dec 2011 and were randomly divided into control group and treatment group. The patients in the control group were giv?en vitamin B1 60mg/d, while the patients in the treatment group were given risperidone 1mg/d. Both groups were treated for 1 month. Positive and negative symptom scale (PANSS) and symptom scale (TESS) were used to evaluate the efficacy and adverse reactions of treatment. Results The PANSS score was significantly lower in the treatment group than in con?trol group at 1, 2, 3 and 4 weeks following treatment(difference between groups:F=48.12 ,P<0.0001;Time difference:F=290.99 ,P<0.0001; Interaction between group and time: F=11.91,P<0.0001 ). After time-adjustment, the PANSS score was significantly lower in the treatment group than in control group at 2, 3 and 4 weeks following treatment. In the course of treatment, the patients in both groups had varying degrees of headache, nausea, weight gain and Beckoning. These side effects were alleviated through symptomatic treatment. Conclusion Risperidone can significantly improve psy?chiatric symptoms in patients with frontotemporal brain contusion with satisfactory safety.

Change rules and correlation between bone mass, bone turnover markers and estrogen levels in different periods of ovariectomized rats / 中国组织工程研究

BACKGROUND:There are so many studies about ovariectomized rats at present, but the research on the change rules of bone mass, bone turnover markers, estrogen levels and their correlation in different periods of ovariectomized rats is rarely reported. OBJECTIVE:To analyze the change rules of bone mass, bone turnover markers, estrogen level and their correlation in different periods of ovariectomized rats. METHODS: Thirty-four 3-month-old female Sprague Dawley rats were randomly divided into three groups: baseline group, ovariectomized group and sham operated group. At the beginning of the experiment, the rats in the baseline group were sacrificed, then rats in the ovariectomized group and sham operated group were executed at 4, 8, 12 weeks postoperative respectively. The bone mineral density, bone mass content, area of different zones of the L1-3 lumbar vertebrae and femurs were detected by dual-energy X-ray absorption method, and meanwhile the serum levels of type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen were determined by ELISA. At last, we analyzed the correlation between body mass, bone mineral densityin vitro, type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen levels and the age of ovariectomized rats. RESULTS AND CONCLUSION: (1) The bone mineral density and bone mass content of the lumbar vertebral and femurs in the ovariectomized group were significantly lower than those in the sham operated group and baseline group at the 4th week after operation (P < 0.05). The bone mineral density and bone mass content in the ovariectomized group were ameliorated obviously at the 8th and 12th weeks compared with those at the 4th week after operation (P < 0.05). The bone mass loss was highest in the L1 and intertrochanteric regions. (2) Serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide in the ovariectomized group were significantly higher than those in the baseline group and sham operated group at the 4th week after operation, but there was no difference at the 8th and 12th weeks. (3) The serum estrogen level in the ovariectomized group was prominently lower than that in the sham operated group and baseline group at the 8th and 12th weeks after operation (P < 0.01 at the 8th week,P < 0.05 at the 12th week). (4) The age was positively correlated with body mass and bone mineral density of the lumbar vertebrae and femursin vitro, while the serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide were negatively correlated with the bone mineral density of the lumbar vertebrae and femurs in vitro (P < 0.01). These results suggested that the bone mass of the lumbar vertebrae and femurs in ovariectomized rats was decreased rapidly firstly, and then rose slowly with time; the bone mass in the L1 and intertrochanteric regions lost seriously; the bone turnover markers showed a significant increase at the beginning of ovariectomy and reduced gradualy to normal condition, while the estrogen level was increased at the first month after ovariectomy and then decreased rapidly. In addition, the body mass, bone turnover markers and estrogen level were associated with the change of bone mass.

Comparison of the effects of gastric gavage and intramuscular injection of prednisone on bone mineral density, skeletal biomechanical properties and bone metabolism in rats / 中国实验动物学报

Objective To compare the effects of gastric gavage and intramuscular injection of prednisone on the bone mineral density, skeletal biomechanical properties and bone metabolism in rats.Methods A total of 45 SPF rats were randomly divided into three groups:normal group, intragastric administration group, and intramuscular injection group.The normal group, as a control group, was administrated with normal saline 2 mL per day, both the intragastric administration group and i.m.injec-tion group received prednisone 0.5 mg/(kg.d) for 12 weeks.All rats were examined for bone mineral density (BMD) and the level of serum β-CTX and PINP.The femoral cortical biomechanical properties ( elastic load, maximal load, rupturing load) were measured by three point bending test.Results After 12 weeks, compared with the normal group, BMD and elastic load, maximal load, and rupturing load of the femur were significantly decreased.Compared with the intragastric gavage group, BMD was significantly decreased, while the elastic load, maximal load, and rupturing load of the femur were not significantly changed in the i.m.injection group (P＜0.05 for all).Compared with the normal group, the level of serum β-CTX was significantly raised (P＜0.05) and the level of serum PINP was significantly decreased (P＜0.05).Compared with the intragastric gavage group, the level of serumβ-CTX was also significantly raised (P＜0.05), the level of serum PINP was significantly decreased (P＜0.05), the bone trabecula and hemopoietic tissue were obviously decreased, while the adipose tissue increased obviously. Conclusions Both intragastric gavage and intramuscular injection of prednisone affect the level of BMD, skeletal biomechanical properties and bone metabolism.However, i.m.injection of prednisone decreases the BMD and bone strength more significantly, leading to a higher bone turnover with increased bone resorption, and leads to osteoporosis earlier.Our results may suggest that oral administration of prednisone is more safe in clinical treatment.

Establishment of a rat model of lumbar nerve root compression via modified unilateral hemilaminectomy approach / 中国实验动物学报

Objective To explore the characteristics and advantages of the rat model of lumbar nerve root com -pression established via modified hemilaminectomy approach .Methods Forty Sprague Dawley ( SD) rats were randomly divided into two groups ( experimental group and control group ) for establishing a rat model of lumbar nerve root compres-sion.Modified hemilaminectomy apprpoach was used in the experimental group , while total laminectomy was applied in the control group.Duration of operation, estimated blood loss, wound healing status, postoperative mortality, motor function of the lower extremities , pathological changes and the gray level of cytoplasmic TNF-αand IL-1 expression were observed to e-valuate the characteristics and reliability of the two operation approaches in establishing a rat model of lumbar nerve root compression.Results Compared with the control group , duration of operation , estimated blood loss , wound healing sta-tus, postoperative mortality were significantly better in the experimental group (P<0.01), whereas there was non-signifi-cant difference between two groups with regard to motor function of the lower extremities , pathological changes and gray lev-el of the cytoplasmic expression of TNF-αand IL-1 between the two groups .Moreover, the soft tissue structural alterations were reduced in the experimental group , which might be helpful for the rat postoperative rehabilitation .Conclusions Modified hemilaminectomy approach is a reliable alternative method for establishing a rat model of lumbar nerve root com -pression, which is beneficial to shorten the operative time , improve wound healing status , reduce estimated blood loss and damages of soft tissue , and decrease mortality rate .Moreover , this modified method is more consistent with the principles of animal ethics .

The Molecular Pathogenesis of Pituitary Adenomas: An Update

Pituitary tumors represent the most common intracranial neoplasms accompanying serious morbidity through mass effects and inappropriate secretion of pituitary hormones. Understanding the etiology of pituitary tumorigenesis will facilitate the development of satisfactory treatment for pituitary adenomas. Although the pathogenesis of pituitary adenomas is largely unknown, considerable evidence indicates that the pituitary tumorigenesis is a complex process involving multiple factors, including genetic and epigenetic changes. This review summarized the recent progress in the study of pituitary tumorigenesis, focusing on the role of tumor suppressor genes, oncogenes and microRNAs.

The Molecular Pathogenesis of Pituitary Adenomas: An Update

Pituitary tumors represent the most common intracranial neoplasms accompanying serious morbidity through mass effects and inappropriate secretion of pituitary hormones. Understanding the etiology of pituitary tumorigenesis will facilitate the development of satisfactory treatment for pituitary adenomas. Although the pathogenesis of pituitary adenomas is largely unknown, considerable evidence indicates that the pituitary tumorigenesis is a complex process involving multiple factors, including genetic and epigenetic changes. This review summarized the recent progress in the study of pituitary tumorigenesis, focusing on the role of tumor suppressor genes, oncogenes and microRNAs.

A marine anthraquinone SZ-685C inhibits cell proliferation and induces apoptosis in GH3 cells / 中华显微外科杂志

Objective To explore the effect of SZ-685C on rat pituitary adenoma GH3 cell line.Methods MTT method evaluated its effect of proliferation and inhibitory concentration 50 (IC5o) on GH3 cells,after treated by 0,2.5,5.0,7.5,10.0,12.5,15.0,17.5,20.0 and 30.0 μmol/L SZ-685C for 48 h,GH3 cells were changed to complete medium for 12 d,crystal violet staining was used to investigate colony formation; microscopy and Hoechst 33342 staining assay observed whether the changes of morphological as the result of apoptosis,then detected cell apoptosis by flow cytometry.Results SZ-685C had a dose-dependent inhibitory effect on GH3 cell proliferation and IC50 was (12.9 ± 0.47) μmol/L,MEF(considered as a control group) had little affect in cell proliferation on the concentration of IC50.Inhibitory effects persisted even on removal of SZ- 685C after 12 d,and SZ-685C blocked colony formation ability of pituitary tumor cells.Apoptotic morphological observation of microscope and Hoechst 33342 staining proved apoptosis during treatment of SZ-685C,flow cytometry detection showed that SZ-685C induces 36.4％ of apoptosis,while only 2.0％ in control group.Conclusion SZ-685C inhibits pituitary tumor cell proliferation and induces apoptosis.SZ-685C can be a new anti-patuitary tumor drug for a further study.

Clinical significance of interleukin-17 producing cell infiltration with TGF-β1 expression in glioma / 中华行为医学与脑科学杂志

Objective To investigate correlation between the amount of interleukin-17 (IL-17) producing cells and the expression of transforming growth factor (31 (TGF-β1) in glioma,and evaluate the clinical values of IL-17 and TGF-pl in predicting the prognosis of glioma. Methods The presence of IL-17 and TGF-pl was measured by immunohistochemistry in 135 human glioma (WHO Ⅰ 18,WHO Ⅱ 45,WHO Ⅲ 53,WHO Ⅳ 19) tissues and 15 normal brain tissues. Results There was no IL-17 positive staining in normal brain tissues. Of 135 glioma specimens showed low TGF-pl expression and 77 (57. 03% ) showed high TGF-pl expression. No TGF-β1 expression was detected in normal brain tissue. Furthermore,TGF-β1 expression was positively correlated with the amount of IL-17 producing cells in glioma tissues ( r=0.285, P<0.01). Compared with the low grade,the levels of IL-17 and TGF-pl positive cells were obviously increased in high grade. The Kaplan-Meier analysis showed that there were significant differences in overall survival (OS) between the IL-17 and TGF-pl high-expression and lowexpression group (P＜0.01, P＜0.05). The 3-year OS rates of IL-17 of high expression and low expression were 33.75% and 76. 36%. Multivariate Cox proportional hazards regression analysis indicated that age,KPS score, IL-17 were independent prognostic factor for OS (P＜0.01). Conclusion Intratumoral IL-17-producing cell density and the expression of TGF-β1 was associated with the malignancy of human glioma.

Inhibitory effects of TNP-470 in combination with BCNU on tumor growth of human glioblastoma xenografts / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10(7)) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm(3), markedly lower than that in the TNP-470 group [(576.10±114.29)mm(3)] and the BCNU group [(473.01±48.04)mm(3)] (both P<0.01). And the xenograft volume in these 3 treatment groups was even much lower than that in the control group [(1512.61±470.25) mm(3)] (all P<0.01). There was no significant difference in the volume of xenografts between the TNP-470 group and the BCNU group (P>0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P<0.01) on the 21th day following treatment. There was no significant difference in the inhibition rate of tumor growth between the TNP-470 group and the BCNU group (P>0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P<0.05). The MVD of xenografts in the 3 treatment groups was markedly reduced as compared with that in the control group (all P<0.05). No significant changes in weights were observed before and after the treatment in each group (all P>0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

Inhibitory effects of TNP-470 in combination with BCNU on tumor growth of human glioblastoma xenografts / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10(7)) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm(3), markedly lower than that in the TNP-470 group [(576.10±114.29)mm(3)] and the BCNU group [(473.01±48.04)mm(3)] (both P0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

Use of dentritic cells pulsed with HLA-A2-restricted MAGE-A1 peptide to generate cytotoxic T lymphocytes against malignant glioma / 华中科技大学学报（医学）（英德文版）

This study developed a novel approach of targeting malignant glioma with pMAGE-A1(278-286)-specific cytotoxic T lymphocytes (CTLs) induced from the peripheral blood mononuclear cells of healthy donors by multiple stimulations with human leukocyte antigen (HLA)-A2-restricted pMAGE-A1(278-286) peptide-pulsed dentritic cells. Cytotoxic assays were performed by the colorimetric CytoTox 96 assay to analyze cytotoxic activity of the induced CTLs against various target cells. The induced CTLs showed approximately 45% specific lysis against T2pMAGE-A1(278-286) (pMAGE-A1(278-286) peptide pulsed T2 cells) and U251 (HLA-A2(+), MAGE-A1(+)) at an effector:target ratio of 40:1, and approximately 5% cytolysis against T2pHIV, A172 (HLA-A2(-), MAGE-A1(+)), K562 and T2 cells without being pulsed with peptide at any effector:target ratio. The specific killing activity of the induced CTLs against T2pMAGE-A1(278-286) and U251 was much more obvious than in any other control group (P<0.05). The cytotoxic activity against the T2pMAGE-A1(278-286) and U251 was significantly eliminated by anti-HLA class I mAb W6/32. These results suggest that pMAGE-A1(278-286) epitope may serve as a surrogate tumor antigen target of specific immunotherapy for treating HLA-A2 patients with malignant glioma.

Knocking-down of Nogo-A gene expression in PC12 cell line by plasmid-based RNAi / 华中科技大学学报（医学）（英德文版）

To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA (short hairpin RNA, or shRNA) was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein (EGFP) gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls (P>0.05), while the expression level in shRNA-transfected group decreased significantly (P<0.05). It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC12 cells.

Knocking-down of Nogo-A Gene Expression in PC12 Cell Line by Plasmid-based RNAi / 华中科技大学学报（医学）（英德文版）

To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA (short hairpin RNA, or shRNA) was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein (EGFP) gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls (P＞0.05), while the expression level in shRNA-transfected group decreased significantly (P＜0.05). It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC 12 cells.

Application of Vacuum Sealing Drainage in the Treatment of Wound Infection of Earthquake Casualty After Amputation / 广州中医药大学学报

Objective To observe the clinical efficacy of vacuum sealing drainage(VSD) in the treatment of large-area wound infection of earthquake casualty after amputation.Methods Seven patients with large-area wound infection of earthquake casualty after amputation received sustained VSD.Results After VSD for 7~26 days,with an average of 13 days,the wound infection in the 7 patients was controlled.No systemic toxicity was found.The result of wound bacterial culture was negative.Of the 7 patients,4 received phase Ⅱ suture,3 received phase Ⅱ skin graft,and all of the grafted skin survived.Conclusion Vacuum sealing drainage exerts certain effect in the treatment of large-area wound infection of earthquake casualty after amputation.