Objective To characterize molecule profiling for hypoxia/hypothermia-conditioned human endothelial cells(EC),and to examine the hypothesis that the interaction between hypoxia/hy- pothermia-conditioned EC and monocytes plays a key role in early hypoxia/hypothermic reperfusion in- jury.Methods Human aortic EC were conditioned in hypoxia/hypothermia chamber at 4℃for 91) rain followed by 4 to 8 h re-warming at 37℃and then analyzed initially by RT-PCR to evaluate RNA transcripts.To examine early cellular interactions,a cellular adhesion study was performed using EC and fluorescence-conjugated DIO labeled purified human monocytes,and CD4~+ T cells as well as PB- MC.Interactions between PKH-26-labeled EC and these cells were carried out to determine EC mem- brane uptake by monocytes.Results RT-PCR analysis demonstrated that hypoxia/hypothermia-con- ditioned EC up-regulated RNA transcripts of inflammatory cytokines,ECE-1 and endothelin,co-stim- ulatory molecules,and adhesion molecules CD54.FACS showed minimal up-regulation of CD54 on hy- poxia-hypothermia-conditioned EC.Low level adhesion of purified CD4~+ T cells to resting and hypoxi- a/hypothermia-conditioned EC was noted.In contrast,high levels of monocyte adhesion to hypoxia/ hypothermia-conditioned EC were observed when compared to resting EC.EC membrane up-take by monocytes but not CD4~+ T cells was revealed by FACS,and the levels of EC membrane up-take were similar to those of the normal EC.Conclusions Hypoxia/hypothermia-conditioned EC demonstrated up-regulation of multiple RNA transcripts associated with inflammatory responses,cellular adhesion and co-stimulation.Human monocyte adhesion to hypoxia/hypothermia-conditioned EC was greater than T cells.Monocytes up-took EC membrane during their interaction.These findings suggest that the initial interaction between monocytes and hypoxia-hypothermia EC,but not T cells,may play a critical role during early reperfusion injury.

Objective To explore whether autoreactive antibody presents in patients with sporadic idiopathic hypoparathyroidism(sIHP).Methods The subjects including 26 patients with sIHP and 112 genealogical members as well as 60 age-and sex-matched healthy controls.Anti-parathyroid antibodies in the sera were assayed by indirect immunofluorescence.The levels of calcium,phosphorus and magnesium as well as intact parathyroid hormone(iPTH)in the sera were tested.Results Positive autoantibodies against parathyroid tissue were demonstrated in 10 patients(38%)with sIHP,significantly higher than that of in genealogical members(10%,?~2=13.42,P

OBJECTIVETo explore the effect of perioperative nutrition support on nutritional condition and complications of the patients with postoperative pancreatic head cancer.

METHODSThirty four patients received perioperative nutrition support, including enteral nutrition and parenteral nutrition (treatment group). Forty eight patients received routine postoperative parenteral nutrition (control group). According to the operative method, these two groups were further divided into two sub-groups: (1) pancreaticoduodenectomy (PD) subgroup, including 13 cases from treatment group, and 24 cases from control group; (2) palliative operation subgroup, including 21 cases from treatment group, and 24 cases from control group. Body weight, total protein (TP), serum albumin (ALB), and the complications after operation were compared.

RESULTSThe concentrations of ALB and TP in the treatment group were significantly higher than those in the control group (P< 0.05). Body weight and TP of the patients received PD in the treatment group were significantly better than those of the control group (P < 0.05).

CONCLUSIONPerioperative nutrition support can improve postoperative nutritional condition and reduce the postoperative complications in patients with pancreatic head cancer.

Adult ; Aged ; Combined Modality Therapy ; Enteral Nutrition ; Female ; Humans ; Male ; Middle Aged ; Nutritional Support ; methods ; Pancreatic Neoplasms ; surgery ; therapy ; Pancreaticoduodenectomy ; Parenteral Nutrition ; Postoperative Complications ; prevention & control

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

AIMTo determine calycosin-7-O-beta-D-glucoside, astragaloside IV and formononetin in Radix Astragali and other relative samples by HPLC-MS.

METHODSHPLC was carried out with Agilent 1100LC/MSD, equipped with Agilent Zorbax SB C18 column (250 mm x 4.6 mm ID, 5 microm) and mass spectrum detector. The mobile phase (CH3CN-H2O) was eluted in gradient mode.

RESULTSThe calibration curves of calycosin-7-O-beta-D-glucoside, astragaloside IV and formononetin were linear in the range of 0.03 - 1.21 microg x mL(-1), 0.35 - 13.86 microg x mL(-1) and 0.38 - 15.22 microg x mL(-1), respectively. These recoveries of samples were from 95% to 105% with RSD less than 1.5%.

CONCLUSIONThe method was employed to analyse 25 samples of Radix Astragali and other relative samples, including Radix Astragali slice, Radix Astragali Preparata, Hedysarum polybotrys Hand. -Mazz, Astragalus ernestii Comb. The contents of three constituents vary greatly because of the species, place of collection and season of harvesting. This method could apply to evaluate the quality of Radix Astragali and it is simple, sensitive and reliable.

Astragalus Plant ; chemistry ; Astragalus membranaceus ; chemistry ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Saponins ; analysis ; Seasons ; Species Specificity ; Spectrometry, Mass, Electrospray Ionization ; methods ; Triterpenes ; analysis

Objective To evaluate the feasibility of different kinds of surgeries to treat hypopharyngeal carcinoma and investigate the methods to conduct one-stage repair after hypopharyngeal carcinoma removal. Methods Thirty nine cases of hypopharyngeal carcinoma were treated from May 2000 to Mar 2007, of which 27 were originated from pyriform sinus, 8 from postcricoid and 4 from posterior pharyngeal wall. One-stage reconstruction for hypopharyngeal and esophageal defect was done with simple and less-damaged hypopharyngeal plasty by local sewing, infrahyoid muscle flap, pectoralis major flap and supplement of esophagus by stomach. Results The defect of 18 cases was repaired by sternohyoid muscle flap, including 13 from pyriform sinus, 4 from root of tongue and 1 from posterior pharyngeal wall. Seven were repaired by substitute of esophagus by stomach, 5 by local sewing, 4 by cervical flap and 2 by pectoralis major myocutaneous flap. All the patients were successfully followed up during the survival days. Half of the cases suffered from slight deglutitory disorder after operation and recovered soon, while 2 cases were still with worse deglutitory function. All the cases were recovered with laryngeal function, and 21 cases were decannulated with normal respiration and voice, and 18 with cannula. Thirteen cases survived for more than 3 years after operation and 5 cases more than 5 years, while 14 cases died within 1 year. Thirteen cases were carried out another operations for local recurrence or metastases of lymph nodes, one of whom experienced 6 operations. The estimated 3-year survival was 43.3% (13/30), and 5-year survival was 37.5%(3/8). Conclusion Hypopharyngeal carcinoma with different site and stage can be treated by different methods, and one-stage reconstruction can be done to repair the defect of pharynx and esophagus with satisfactory functional restoration and prognosis.

BACKGROUNDThe aim of this study was to assess the performance of apparent diffusion coefficient (ADC) measurement obtained with diffusion-weighted magnetic resonance imaging (DW-MRI) to distinguish renal cell carcinomas (RCCs) from small benign solid renal tumors (≤ 4 cm).

METHODSIn this cross-sectional study, 49 consecutive patients with histopathologically confirmed small solid renal tumors, and seven healthy volunteers were imaged using nonenhanced MRI and DW-MRI. The ADC map was calculated using the b values of 0, 50, 400, and 600 s/mm 2 and values compared via the Kruskal-Wallis and Mann-Whitney tests. The utility of ADC for differentiating RCCs and benign lesions was assessed using a receiver operating characteristic curve. Multiple nonenhanced MRI features were analyzed by Logistic regression.

RESULTSThe tumors consisted of 33 cases of clear-cell RCCs (ccRCCs) and 16 cases of benign tumors, including 14 cases of minimal fat angiomyolipomas and 2 cases of oncocytomas. The ADCs showed significant differences among benign tumors ([0.90 ± 0.52] × 10-3 mm 2 /s), ccRCCs ([1.53 ± 0.31] × 10-3 mm 2 /s) and the normal renal parenchyma ([2.22 ± 0.12] × 10-3 mm 2 /s) (P < 0.001). Moreover, there was statistically significant difference between high and low-grade ccRCCs (P = 0.004). Using a cut-off ADC of 1.36 × 10-3 mm 2 /s, DW-MRI resulted in an area under the curve (AUC), sensitivity, and specificity equal to 0.839, 75.8%, and 87.5%, respectively. Nonenhanced MRI alone and the combination of imaging methods led to an AUC, sensitivity and specificity equal to 0.919, 93.9%, and 81.2%, 0.998, 97%, and 100%, respectively. The Logistic regression showed that the location of the center of the tumor (inside the contour of the kidney) and appearance of stiff blood vessel were significantly helpful for diagnosing ccRCCs.

CONCLUSIONSDW-MRI has potential in distinguishing ccRCCs from benign lesions in human small solid renal tumors (≤ 4 cm), and in increasing the accuracy for diagnosing ccRCCs when combined with nonenhanced MRI.

Adult ; Aged ; Carcinoma, Renal Cell ; diagnosis ; Cross-Sectional Studies ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Kidney Neoplasms ; diagnosis ; Male ; Middle Aged ; Young Adult

PURPOSETo develop a novel injectable strontium-containing calcium phosphate cement with collagen.

METHODSA novel calcium phosphate bone cement (CPC) was prepared with the addition of strontium element, collagenl, and modified starch; the injectability, solidification time, microstructure, phase composition, compressive strength, anti-collapsibility and histological properties of material were evaluated.

RESULTSThe results showed that the material could be injected with an excellent performance; the modified starch significantly improved the anti-washout property of cement; with the liquid to solid ratio of 0.3, the largest compressive strength of cement was obtained (48.0 MPa ± 2.3 MPa); histological examination of repair tissue showed that the bone was repaired after 16 weeks; the degradation of cement was consistent with the new bone growth.

CONCLUSIONA novel injectable collagen-strontium-containing CPC with excellent compressive strength and suitable setting time was prepared, with addition of modified starch. The CPC showed a good anti-washout property and the degradation time of the cement met with the new bone growing. This material is supposed to be used in orthopedic and maxillofacial surgery for bone defects.

Animals ; Bone Cements ; chemistry ; therapeutic use ; Calcium Phosphates ; chemistry ; Collagen ; chemistry ; Compressive Strength ; Histocompatibility Testing ; Injections ; Rabbits ; Strontium ; chemistry

This study was purposed to culture murine compact bone-derived mesenchymal stem cell (MSC) and analyze the immunological and trilineage differentiation potential. Tibia and femur were extracted. Bone marrow cells were flushed out and compact bone fragments were digested with collagenase. The digested cells were cultured in 6-well plates. The immunophenotype, immunosuppressive function and trilineage differentiation potential were analysed by flow cytometry, mixed lympocyte reaction and Oil red O, von Kossa and alcian blue straining, respectively. The results indicated that the pure compact bone MSC could be isolated with in 3 weeks. The resulting MSC had trilineage differentiation potential and immunosuppressive effect on mixed lymphocyte reaction. The count per minute (CPM) value in control group of BALB/c T cells cocultured with irradiated C57BL/6 T cells was (2.56 ± 0.31) × 10(4), while CPM values of mixed lymphocyte cocultured with C57BL/6 compact bone MSC at ratios of 100:1 and 10:1 were (0.47 ± 0.12) × 10(4) and (0.28 ± 0.09) × 10(4). The CPM value of control group was higher than those of MSC cocultured group (P < 0.001). Compact bone-MSC had an immunosuppressive effect on mixed lymphocyte reaction in a dose dependent manner. It is concluded that murine compact bone has rich MSC and the primary MSC is contaminated with less hematopoietic cells. Murine compact bone-MSC have immunosuppressive effect on mixed lymphocyte reaction and trilineage differentiation potential. Compact bone-MSC have promising experimental study value.
Animals ; Bone Marrow Cells ; cytology ; immunology ; Bone and Bones ; cytology ; Cells, Cultured ; Female ; Immunophenotyping ; Lymphocyte Culture Test, Mixed ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

To improve the after-sales service, a survey aimed at the after-serveis of 3 kinds of medical equipment is applied among 68 hospitals in Shanghai Area in 2008.The Stat. and analysis results are showed in the paper, which will certainly channel off suppliers to set up a harmonious market together.
China ; Consumer Behavior ; statistics & numerical data ; Data Collection ; Product Surveillance, Postmarketing