Objective To reverse P-glycoprotein-mediated muhiple drug resistance using RNA interference(RNAi)in cultured rat astrocytes.Methods Astroeytes overexpressing P-glyeoprotein induced by coriaria lactone were transfected with the short-hairpin RNA expression vector-p-SIREN shuttle designed to target Mdrl mRNA.The mRNA level of Mdrl gene was evaluated by real-time PCR;the P-glycoprotein was examined by immunocytochemistry and image analysis,meanwhile the rhodamine efflux was assessed by flow cytometry.Results The astrocyte model overexpressing P-glycoprotein were established and successfully transfected with the short-hairpin RNA expression vector-p-SIREN shuttle.The mRNA level of Mdrl gene was knocked down by 67.70%(P

Objective:To analyze the characteristics and related risk factors of femoral supracondylar fractures combined with meniscus and ligament injuries.Methods:A retrospective case series analysis was performed on clinical data of 100 patients with femoral supracondylar fractures combined with ipsilateral meniscal and?ligamentous knee?injuries?admitted to Tianjin Hospital from January 2016 to December 2020. There were 55 males and 45 females，with age of 22-78 years［（38.5 ± 3.3）years］. Causes of injury included traffic accidents in 45 patients，falls in 45 and others in 10. According to AO classification，type A1 was in 56 patients，A2 in 33 and A3 in 11. Sixty of these patients presented fractures on the left side and forty on the right side. All patients accepted MRI examination to estimate meniscal and ligamentous knee injuries concerning its prevalence，locations and severity. Locations of injury included the medial meniscus（MM），lateral meniscus（LM），anterior cruciate ligament（ACL），posterior cruciate ligament（PCL），medial collateral ligament（MCL）and lateral collateral ligament（LCL）. Meniscal knee injuries were measured using Stoller four-level method，and ligamentous knee injuries by Ruiz three-level method. The patients were subdivided based on age，sex，causes of injury，injury side and fracture AO classification to measure the prevalence of meniscal and?ligamentous injuries，and to identify the factors related to injuries using logistic regression analysis.Results:Seventy-six patients（76.0%）were complicated with meniscal or ligamentous injuries in 151 different parts. Twenty-nine patients（29.0%）had both meniscal and ligamentous injuries. Fifty-six patients had meniscal injuries，including 11 with bilateral meniscus injuries. MM injuries were noted in 32 patients（32.0%），with severity of grade Ⅰ in 19 patients，grade Ⅱ in 12 and grade Ⅲ in 1. LM injuries were found in 35 patients（35.0%），with severity of grade Ⅰ in 19 patients，grade Ⅱ in 14 and grade Ⅲ in 2. Forty-nine patients had ligamentous injuries，among which 22 patients were with single ligamentous injuries，19 with double ligamentous injuries and 8 with triple ligamentous injuries. ACL injuries were found in 33 patients（33.0%），with severity of grade Ⅰ in 22 patients，grade Ⅱ in 10 and grade Ⅲ in 1. PCL injuries occurred in 4 patients（4.0%），with severity of grade Ⅰ in 3 patients and grade Ⅱ in 1. MCL injuries occurred in 28 patients（28.0%），with severity of grade Ⅰ in 18 patients，grade Ⅱ in 9 and grade Ⅲ in 1. LCL injuries occurred in 19 patients，with severity of grade Ⅰ in 12 patients，grade Ⅱ in 5 and grade Ⅲ in 2. There were significant differences in prevalence of meniscal injuries among subgroups of different gender，causes of injury and fracture types（ P < 0.05）.Logistic regression analysis suggested that gender and fracture types showed significant correlation with meniscal injuries，especially factors of males（ P < 0.05）and type A3 fractures（ P < 0.01）. There were significant differences in prevalence of ligamentous injuries among subgroups of different causes of injury and fracture types（ P < 0.05）. Logistic regression analysis showed that causes of injury and fracture types were significantly related to ligamentous injuries，especially factors of traffic accidents（ P < 0.01）and type A3 fractures（ P < 0.01）. Conclusions:Ipsilateral meniscal or ligamentous knee injuries have a high incidence in patients with supracondylar femoral fractures. There exist multiple-site injuries，particularly in ligamentous injuries. Males，type A3 fractures and traffic injuries are risk factors，which needs to be paid much attention in clinical diagnosis and treatment.

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

BACKGROUND:The laparoscopy is superior to open surgery for being less invasive, inducing mild stress reaction and allowing quick recovery after operation, however the effects of laparoscopy on perioperative serum cytokine levels are controversial, and only a few studies discuss these effects among pediatric patients.OBJECTIVE: To compare the changes in perioperative cytokine levels and their clinical significance in pediatric patients undergoing laparoscopy and open surgery.DESrGN: Non-randomized concurrent controlled observation.SETTTNG: Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA.PARTICT PANTS: From May 2004 to December 2006, 135 pediatric patients for elective operation were recruited from Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA. Sixty-five patients were scheduled for laparoscopic surgery while the remaining 70 patients for open surgery.METHODS: Serum levels of interleukin-1 beta (IL-1β), IL-6, IL-10 and tumor necrosis factor-alpha (TNF-α) were measured at 24 hours before operation, and 3, 24, 48 hours after operation respectively. Duration of hospitalization time of all the children was also recorded.MAIN OUTCOME MEASURES: Levels of IL-1β, IL-6, IL-10 and TNF-α of all the patients were measured 24 hours preoperatively, and 3, 24, 48 hours postoperatively.RESULTS: All the 135 cases were included for statistical analysis. ①There were no significant perioperative changes in cytokine levels after laparoscopic surgery (P ＞ 0.05). In the open surgery group, IL-1β and IL-6 levels increased significantly at 3 and 24 hours after operation (P ＜ 0.05), and normalized within 48 hours postoperatively. No significant perioperative differences were found in IL-10 and TNF-α levels (P ＞ 0.05). The levels of IL-1β and IL-6 were significantly higher in the open surgery group than in the laparoscopic surgery group (P ＜ 0.05). ②Duration of hospitalization was shorter in the laparoscopic surgery group than in the open surgery group [(3.5±1.0), (7.5+1.5) days, P＜ 0.05].CONCLUSTON: Pediatric patients undergoing laparoscopic surgery had less perioperative changes in cytokine levels and quicker recovery.

BACKGROUNDUsing magnetic resonance imaging, diagnosis of malignant meningioma from benign meningioma with atypical features is uncertain. We evaluated the value of lipid signal in differentiating intracranial meningiomas.

METHODS1H-magnetic resonance spectroscopy (MRS) using a point resolved spectroscopy (TR/TE 1000/144 ms) sequences were performed on 34 patients on a 3.0 T scanner. Lipid peak located at 1.3 ppm was evaluated. MRS data from these tumours were compared with histopathological findings (including hematoxylin and eosin staining and KP-1 staining).

RESULTSTwenty-nine meningiomas were histologically benign (eleven meningothelial, thirteen fibrous, four transitional and one microcystic), three were atypical, and two were anaplastic. Lipid signal was detected in ten cases: two anaplastic, three atypical, two fibrous and three meningothelial meningiomas. All voxels with lipid peak in the spectrum from the tumour were evaluated. With creatinine peak in the normal white matter chosen as internal standard, lipid/creatinine ratios of anaplastic, atypical and benign meningiomas were 0.844 +/- 0.027 (range from 0.725 to 0.994), 0.465 +/- 0.023 (range from 0.239 to 0.724), and 0.373 +/- 0.016 (range from 0.172 to 0.571) respectively. Highly significant differences were noted between anaplastic and the other two subtypes. Patchy necrosis was observed in anaplastic meningioma, while focal necrosis was noted in atypical meningioma with HE stain. However, no necrosis was found in benign group. KP-1 stain demonstrated histocytes containing lipids in the necrotic region of anaplastic and atypical meningioma.

CONCLUSIONThe lipid signal at 1.3 ppm is a useful marker in evaluating the malignancy of intracranial meningiomas, especially in the differential diagnosis of anaplastic meningioma.

Adult ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Magnetic Resonance Spectroscopy ; methods ; Male ; Meningeal Neoplasms ; diagnosis ; Meningioma ; diagnosis ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity

Objective: To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD). Methods: Recipient BALB/c mice received 8 × 10⁶ bone marrow cells with 8×10⁶ spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4⁺ T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry. Results: Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ²=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro. Conclusions Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.

OBJECTIVETo evaluate the effect of comprehensive control strategy of schistosomiasis with emphasis on infection source control in Anhui province.

METHODSForty endemic villages in Guichi district, Chizhou city, Anhui province were selected as national pilot villages in the years from 2006 to 2008, and another 10 provincial pilot villages were respectively selected from 10 highly endemic villages in 7 cities in 2007. The comprehensive infection source control measures, including "replace cattle with machines", "raise livestock in pens", "improve the sanitary toilets", "supply safe water " and so on were carried out among the above pilot villages. At the end of 2008, 13 national pilot villages and 6 provincial pilot villages were selected to investigate the popularity of schistosomiasis, and the effect of the comprehensive control strategy in those villages were compared.

RESULTSAfter implementing the comprehensive control strategy, the infectious rate of schistosomiasis in national pilot villages decreased from 4.57% (487/10 659) to 1.76% (147/8370), with the reduction rate at 61.49%, whose difference showed statistical significance (χ(2) = 115.16, P < 0.01); and the density of infected snails decreased from 0.0067/0.1 m(2) to 0.0008/0.1 m(2), the infectious rate of snails decreased from 0.28% to 0.04%, whose reduction rates were 88.06% and 85.71% respectively. While as to the provincial pilot villages, the infectious rate of schistosomiasis decreased from 1.27% (54/4254) to 0.21% (14/6592), with the reduction rate at 83.46%, whose difference showed statistical significance (χ(2) = 94.57, P < 0.01); and the density of infected snails decreased from 0.0025/0.1 m(2) to 0.0003/0.1 m(2), the infection rate of snails decreased from 0.13% to 0.05%, whose reduction rates were 88.00% and 61.54% respectively.

CONCLUSIONThe comprehensive control strategy with emphasis on infection source control implemented in marshland and lake regions can effectively control the transmission of schistosomiasis.

Animals ; China ; epidemiology ; Disease Reservoirs ; parasitology ; Humans ; Infection Control ; methods ; Livestock ; Pilot Projects ; Rural Health ; Schistosomiasis ; epidemiology ; parasitology ; prevention & control ; Snails ; parasitology

OBJECTIVETo clone and identify gastric cancer-related genes and explore the possible pathogenic mechanism of gastric cancer.

METHODSThe differentially expressed cDNA bands were isolated by fluorescent mRNA differential display in gastric cancer specimens, matched normal gastric mucosa and premalignant lesions. The motility-related protein-1 (MRP-1/CD9) gene was one of the down-regulated genes. MRP-1/CD9 gene expression in different kinds of gastric tissue was analyzed by Northern blot and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSMRP-1/CD9 expression was down-regulated in all gastric cancer tissues. Northern blot analysis confirmed this differential expression. RT-PCR analysis showed that the MRP-1/CD9 gene expression was much lower in gastric cancers(0.31+/-0.18) than in the matched normal gastric tissue (0.49+/-0.24) and in the premalignant lesions (0.47+/-0.18) (P<0.05), and its expression in intestinal type gastric cancer (0.38+/-0.16) was higher than that in diffuse type gastric cancer (0.22+/-0.17) (P<0.05).

CONCLUSIONThe MRP-1/CD9 gene expression was down-regulated in gastric cancer, its expression was probably related to the carcinogenesis and histology types of gastric cancer.

Antigens, CD ; genetics ; Blotting, Northern ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Membrane Glycoproteins ; genetics ; Precancerous Conditions ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; Tetraspanin-29

OBJECTIVETo screen the molecular markers for refractory anemia with excess blasts in transformation (RAEB) in myelodysplastic syndromes (MDS) by serum proteome profiling.

METHODSThe serum protein were isolated from patients with RAEB, acute myeloid leukemia or normal subjects by 2-dimensional electrophoresis (2-DE), and the electrophoresis gels were obtained to identify the differentially reacting protein spots. The replica gels of the differentially reacting proteins were analyzed to locate the matching protein spots, which were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) and database searching.

RESULTSSeven differentially expressed proteins in RAEB were found by 2-DE. Of the 7 proteins, 4 were identified by MALDI-TOF-MS to have significantly differential expression in RAEB, including dipeptidyl peptidase (DPP/CD26), polymerase (DNA directed) kappa, PRO2044 and an albumin-like protein.

CONCLUSION2-DE-based serum proteome profiling helps identify serum proteomic biomarkers related to MDS. DDP/CD26 has increased expression in the serum in RAEB subtype MDS, suggesting its possible role in advanced MDS.

Anemia, Refractory, with Excess of Blasts ; blood ; genetics ; Bone Marrow ; pathology ; DNA-Directed DNA Polymerase ; blood ; Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ; blood ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; classification ; genetics ; Proteomics

OBJECTIVETo study the clinical effects and application value of intraoperative CT in treatment of severe scoliosis with posterior total pedicle screws.

METHODSThirty-two cases of severe scoliosis were retrospectively analysed in our hospital from June 2009 to June 2011,which were treated by posterior total pedicle screws with intraoperative CT including 12 males and 20 females with an average age of 16.8 years ranging from 10 to 38 years. There were 19 cases combined with thoracic kyphosis among 32 cases. Multiple planar reconstruction technology of intraoperative CT was applied to assess screw position. The numbers (rates) of pedicle screws were calculated and evaluated as different grades in upper thoracic vertebra (T1-T4) ,middle thoracic vertebra (T5-T8), lower thoracic vertebra (T9-T12) and lumbar vertebra. The pedicle screws of 2 grade and 3 grade were defined as malpositioned screws. Times of applicating intraoperative CT were calculated. Cobb angle of all cases and kyphosis angle of the cases combined with thoracic kyphosis were measured before and after surgery. Scoliosis correction rates and kyphosis correction rates were calculated.

RESULTSThere were 686 pedicle screws placed in thoracolumbar of 32 patients (including 544 thoracic pedicle screws,142 lumbar pedicle screws) and 14 patients underwent osteotomy. The rate of malpositioned screws in thoracolumbar was 7.3% by evaluating with intraoperative CT,and it respectively was 5.6%,11.1%, 6.7% and 4.3% in upper thoracic vertebra, middle thoracic vertebra,lower thoracic vertebra and lumbar vertebra. The malpositioned screws were amended in surgery. The mean times of intraoperative CT was 2.6 times (ranged from 2 times to 4 times). The mean preoperative Cobb angle was 95 degrees (ranged from 78 degrees to 123 degrees) and the mean postoperative Cobb angle was 340 (ranged from 19 degrees to 53 degrees). The mean correction rate of Cobb angle was 64%. The mean preoperative kyphosis angle of the patients combined with thoracic kyphosis was 69 degrees (ranged from 46 degrees to 82 degrees) and the mean postoperative kyphosis angle was 32 degrees (ranged from 22 degrees to 45 degrees). The mean correction rate of kyphosis angle was 54%. Four patients suffered cerebrospinal fluid leak after surgery. No infection, vascular lesion and nervous lesion were found. All patients had an average 18-month follow-up (ranged from 12 to 26 months). No broken nails, broken rods and pseudarthrosis were founded.

CONCLUSIONApplication of in traoperative CT in severe scoliosis with posterior total pedicle screws can detect and amend malpositioned screws timely in surgery, to avoid secondary surgery for malpositioned screws and protect the safety of surgery. The effects of surgery is satisfactory.

Adolescent ; Adult ; Bone Screws ; Child ; Female ; Humans ; Male ; Monitoring, Intraoperative ; Retrospective Studies ; Scoliosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; methods