Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Malathion ; metabolism ; urine

Human platelets have distinct characters when preserved by different methods. A efficient flow cytometric assay for different preserved platelets expression of CD62p and phosphatidylserine(PS) is in dire need. Efficient flow cytometric assay for CD62p and PS expressed by preserved platelets was established and the major conditions were optimized. The platelets need not to be washed to wipe off plasma and can be labelled diredtly during the sample preparation. It is efficient for flow cytometric analysis when fresh platelet riched plasma (FPRP) was set as negative control, thrombin actived FPRP, and liquid nitrogen treated FPRP were set as positive control respectively. Gly-Pro-Arg-Pro acetate salt (GPRP) was applied to prevent platelets aggregation and fibrin formation, stabilize platelets and minimize the artificial platelets activation. This is also the key to conquer difficulty of flow cytometric quantitive analysis when platelet, Ca(2+) and plasma coexist. This flow cytometric method is specially suitable for the multi-parameter assay including PS expression for cryopreserved platelets. Minimal sample manipulation, no fixation, and GPRP application resulted in minor artifacts and good sample stability. Results suggested, this flow cytometric assay for preserved platelets is simple and efficient. In addition, the author prepared four different methods treated platelets that can be easily distinguished through this flow cytometric assay. It not only makes sure the practicability of this flow cytometric assay, but also suggests the value of the treated platelets applied in preserved platelets flow cytometric ass
Blood Platelets ; metabolism ; Flow Cytometry ; methods ; Humans ; P-Selectin ; biosynthesis ; Phosphatidylserines ; analysis ; Reproducibility of Results ; Tissue Preservation

BACKGROUND:Alveolar bone remodeling and sustained absorption due to tooth extraction seriously affect the implanting conditions and morphology of hard and soft tissue in implant zone. OBJECTIVE:To evaluate the effect of nano-hydroxyapatite/polycaprolactone electrospinning scaffolds to improve the osteogenic effect of bone defects around immediate implants. METHODS:Tissue-engineered bone was prepared by combining canine bone marrow mesenchymal stem cels with nano-hydroxyapatite/polycaprolactone electrospinning scaffold. Bilateral mandibular second premolars from six dogs were extracted mandibular second premolar, and an immediate implant was placed in the mesial fossa of the mandibular second premolar. Three-wal bone defects was made buccaly using titanium nails, then tissue-engineered bone and Bio-Oss bone powders were implanted bilateraly covered by colagen membranes (Bio-Gide). Imageology examination was performed to measure bone gray levels immediately, 4, 8, 12 weeks after surgery. After 12 weeks, the mandible was removed completely, toluidine blue staining was used for observation of microstructure, new bone formation, bone morphology and implant osseointegration. RESULTS AND CONCLUSION: Between the two groups, there was no difference in bone mineral density at each time point after surgery, indicating that the effects of the two materials to promote bone regeneration process are basicaly the same. After implantation, the dense lamelar bone formed in the bone defect region of tissue-engineered bone group, mature bone cels, Haversian canal, and implant osseointegration were visible. While, in the Bio-Oss group, the lamelar bone was dense, a smal amount of Bio-Oss particles distributed within new bone tissues, fewer bone cels were found, a part of Haversian canal was shown to have blood capilaries, and new bone was in close conjunction with the implant. These findings indicate that the nano-hydroxyapatite/polycaprolactone electrospinning scaffold combined with bone marrow mesenchymal stem cels and Bio-Gide colagen membrane can promote the regeneration of alveolar bone around the implant.

Objective To investigate the clinical value of spinal nonfusion interspinous Coflex system for the degenerative lumbar spinal stenosis.Methods From March 2008 to August 2009,26 patients with degenerative lumbar spinal stenosis were treated by decompression with posterior spinous process and interspinous implant Coflex folder method,including 11 males and 15 females,with the mean age of 65.4years(range,45-78 years).L3,4 segment in 7 cases,L4,5 segment in 13 cases,concomitant L3,4,L4,5 segment stenosis in 6 cases.Patients were scanned by MRI and CT confirmed L3,4 and(or)L4,5 segments of the ligamentum flavum thickening,proliferation of small-joint bone,merger disc herniation causing central tube and lateral fossae stricture,nerve root or coccygeal plexus compression.Lumbar anteroposterior,lateral and flexion-extension X-ray films preoperatively,and at follow-up were used to measure the following parameters by eFilm and CAD software:the anterior and posterior disc space height,the range of motion at surgical level,and the segment of the spinal canal area of responsibility.Postoperative standard Japanese Orthopaedic Association(JOA)score for preoperative and postoperative scores.Results Patients were followed up 12-24months.The pain relieved in all patients.The JOA scores improved from(15.46 ±4.30)preoperatively to (24.50±1.58)postoperatively,the vertebral canal area of the affected segment was(218.4±16.2)mm2 before the operation,(264.6±9.9)mm2 after the operation.Single segment anterior disc space height did not change significantly,the difference was not statistically significant.The disc space posterior height increased compared with the preoperative height,and gradually decreased with time.No patients suffered Coflex loosening,fracture and emerge.Conclusion The treatment of lumbar spinal stenosis with Coflex system has satisfactory effect in minimal invasiveness and high security,which provides a safe and effective therapy for degenerative lumbar spinal stenosis.

Objective To study the effect and related mechanism of celecoxib on tumor necrosis factorrelated apoptosis-inducing ligand(TRAIL) induced apoptosis of medullary thyroid cancer TT cell line.Methods MTT assay was used to measure the growth inhibition induced by TRAIL and celecoxib alone and their combination.TT cell cycle distribution was analyzed by flowcytometry.Hochest33258 staining and DNA ladder was used to detect the apoptosis of drug combination on TT cells.Western blot was used detect the protein change of cyclin A,Cdk2,caspase-8,c-FLIP,and RIP.Results ①MTT showed the growth inhibition ratio of TT cell intervened by the combination of TRAIL and celecoxib was 47.53％ ± 1.34％,which was much higher than that intervened by TRAIL(7.75 ％ ± 3.84％)and celecoxib alone.The differences had statistical significance (t test,F =5.234,P ＜0.01);②PI detection found the cells' number in G0/G1 phase in celecoxib group and combination group were increased compared to that in control group and TRAIL group(F =242.694,P ＜ 0.01);③Western blot indicated the expression of Cyclin A and Cdk2 were down regulated,there was no statistic significance;④ The apoptosis morph in nuclus was detected by Hochest33258 staining and showed the karyopycnosis and muclear fragmentation were increased in combination group with the apoptosis rate 24.23％ ± 2.91％,which was much higher than that in TRAIL(5.86％ ± 1.41％) and celecoxib(20％ ± 1.24％) (t test,F =1.824,P ＜0.01),the difference has statistic significance;⑤Western blot illustrated the active schizolysis of casplase-8 was higher and the expression of c-FLIP and RIP was down regulated in combination group.Conclusion celecoxib plays a positive effect on TRAIL-reduced apoptosis of medullary thyroid cancer TT cell line,which may due to the cell cycle arrest at G0/G1 phase,down-regulation of c-FLIP and RIP and subsequent activation of caspase-8.

Objective: To investigate the relationship between chemosensitivity to L-OHP and expressions of multidrug resistance (MDR) associated factors in lymph node metastases (LNMs) of gastric carcinoma. Methods: The chemosensitivity to L-OHP was measured by MIT assay, and the expressions of P-gp, GST-π, P53, Survivin and Bcl-2 were determined by immunohistochemistry in 54 paired primary tumor (PT) and LNMs of gastric carcinoma. Results: The inhibition rates of LNMs cells for L-OHP were lower than those of PT (P<0.05). The expressions of P-gp, GST-π and Bcl-2 were higher in LNMs than in PT (P<0.05), and no signifi-cant difference was found in the expression of P53 and Survivin between LNMs and PT (P>0.05). Positive cor-relations among P-gp, P53 and Bcl-2 were found in PT and LNMs (r=0.3424, 0.7123, 0.4548, P<0.05). There was no significant difference in the expression of GST-π and Survivin between PT and LNMs (P>0.05). There was statistically negative correlation between inhibition rates and expression of P-gp, GST-π, and Survivin in PT (P<0.05). In LNMs, only Survivin was negatively correlated with inhibition rates of L-OHP (P<0.05). Conclu-sion: The LNMs of gastric carcinoma are heterogeneous with PT in respect to chemosensitivity to L-OHP and expression of multidrug resistance associated factors. The main factors that affect chemosensitivity to L-OHP are also significantly different between PT and LNMs. Effective adjuvant chemotherapy after surgery and re-version to multidrug resistance (MDR) of gastric carcinoma depend on targeting the metastatic lesions of gas-tric carcinoma.

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

The present study aims to investigate the effects of soluble endoglin (sEng) on invasive ability of cultured cytotrophoblasts of first trimester of pregnancy. Cytotrophoblasts of normal 6 to 8-week pregnancy were cultured by trypsin digestion method, and were incubated with cell culture medium without (control group) and with 10 μg/L sEng (sEng group), respectively for 24 h. The invasive ability was determined by transwell invasion assay, and expressions of MMP-2, MMP-9 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively. The results showed that the invasive ability of cytotrophoblasts in sEng group was lower than that in control group (P < 0.05). Compared with control group, the expressions of MMP-2 and MMP-9 mRNA and protein of cytotrophoblasts were significantly lower (P < 0.05). In conclusion, sEng may participate in the genesis of preeclampsia by affecting the invasive ability of cytotrophoblasts through regulation of the expression of MMP-2 and MMP-9.
Antigens, CD ; pharmacology ; Cell Movement ; drug effects ; physiology ; Cells, Cultured ; Endoglin ; Female ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Placentation ; physiology ; Pre-Eclampsia ; physiopathology ; Pregnancy ; Pregnancy Trimester, First ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cell Surface ; Trophoblasts ; cytology

Objective To investigate the status of medical institution sewage treatment in Qingpu District of Shanghai, and to make corresponding measures in improving sewage management in medical institutions. Methods Questionnaire survey was conducted on selected 95 medical institutions in Qingpu District to investigate sewage treatment status and sewage samples were collected from 30 medical institutions with 20 beds or above to test their sanitary quality. Results Operating rate of sewage handling equipment was 98.9%, the proportion of medical institutions using chlorine disinfectant was 87.4%, the total qualified rate of residual chlorine was 66.7%, the total qualified rate of fecal coliform number was 70.0%, and overall qualified rate was 60.0%. Conclusion The qualified rate of medical institutions sewage in Qingpu District is not optimistic enough.It is necessary to strengthen sewage personnel training and health supervision to ensure the safety of medical institution sewage.

In order to explore the factors that affect CD62p expression in platelet during the whole course of cryopreservated platelets preparation, CD62p expression of platelet was evaluated by flow cytometry assay. The whole course of cryopreservated platelets preparation in order included whole blood collection, centrifugation for fresh platelet-rich plasma preparation, addition of dimethyl sulfoxide, and freeze in -80 degrees C refrigerator and thaw in 38 degrees C water bath. Result showed that the CD62p expressed slowly from whole blood collection to addition of dimethyl sulfoxide, but expressed abruptly after freeze and thaw and it occupied 82 per cent of the whole expression. It was concluded that the whole blood collection, centrifugation for fresh platelet enriched plasma preparation and addition of dimethyl sulfoxide were optimized in the whole course, but the damage to platelets in the whole course of cryopreservation could not be avoided. It suggests that improved techniques are needed to reduce the damage to cryopreservated platelets.
Blood Platelets ; metabolism ; Blood Preservation ; methods ; standards ; Cryopreservation ; methods ; standards ; Flow Cytometry ; Humans ; P-Selectin ; biosynthesis