BACKGROUND: Sinomenine, an alkaloid monomer extracted from Chinese medicinal herb sinomenium acutum,possesses potent anti-inflammatory, analgesic and immunoinhibitory pharmacological activities. Sinomenine has certain therapeutic effect on rheumatoid arthritis and has been widely applied in the clinic. Dendritic cell (DC) is the known antigen presenting cell with the strongest functions in the body.OBJECTIVE: To observe the effect of different doses of sinomenine on CD80 and CD 86 expression and extracellular interleukin-12 secretion in DCs.DESIGN: A controlled repeated measuring study based on the cells.SETTTNG: Department of Pharmacology, Guangdong Medical College.MATERIALS: The experiment was carried out in the Institute of Immunology, Third Military Medical University of Chinese PLA between September 2004 and May 2005. The peripheral blood of healthy donors was supplied by the blood bank of the Southwest Hospital. RPMI1640 medium was purchased from Hyclone Company. Fluorescein isothiocyanate(FITC)-conjugated mouse monoclonal antibodies against CD80 and CD86 and FITC-conjugated mouse IgG1 were purchased from Bioscience Company. Recombinant human interleukin-4 (rhlL-4), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and tumor necrosis factor-α (TNF-α) were purchased from PharMingen Company. Sinomenine was supplied by Zhengqing Pharmacy Company in Hunan. ELISA kit specific for IL-12 was purchased from Diaclone Company.METHODS: Peripheral blood was collected from healthy volunteers. 1 ×106 U/L recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and 5×105 U/Lrecombinant human interleukin-4 (rhlL-4)were added to the peripheral blood. After 7 days, DCs were harvested. ①The DCs were counted and cultured at 2×108 L 1 in fresh medium supplemented with TNF-α and different doses of sinomenine(3, 10, 30 mg/L). After 48 hours, 2×105 cells were collected and washed twice with phosphate buffer solution (PBS). Then the expressions of CD80 and CD86 were analyzed using 20 μL FITC-conjugated mouse monoclone antibodies against CD80 or CD86.FITC-conjugated mouse IgG1 was used as control at 4 ℃ away from light for 30 minutes. After staining, cells were washed twice with PBS and fixed in 10 g/L paraformaldehyde. The samples were analyzed on a flow cytometry. ②The DCs were counted and cultured at 2 ×108 L-1 in fresh medium supplemented with TNF-α and different doses of sinomenine (3, 10, 30 mg/L). After 48 hours, the supernatants were collected and the concentration of interleukin-12 (IL-12) was determined in a sandwich ELISA using kit specific for IL-12 according to the manufacture's instruction.MAIN OUTCOME MEASURES: ①Effect of different doses of sinomenine on the expressions of CD80 and CD86 on DCs. ②Effect of different doses of sinomenine on the level of IL-12 secreted by DCs.RESULTS : ① Expressions of CD80 and CD86 on DCs: There were no significant differences in the percentage of cells positive and fluorescence intensity for CD80 and CD86 on DC between different doses of sinomenine groups and control groups (P ＞ 0.05). ② IL-12 level secreted by DCs: From ELISA measurement, we found that IL-12 level of the sinomenine 3 mg/L, 10 mg/L and 30 mg/L groups was (863.1±33.7), (668.8±31.9), (512.6±29.6) ng/L, respectively,which was higher than that of control group [ (922.2±36.6),P ＜ 0.05-0.01]. The level of IL-12 in the supernatants of DCs co-cultured with sinomenine was significantly lower compared with control, which was dose-dependent.CONCLUSTON: Sinomenine can inhibit the level of IL-12 secreted by DCs in dose-dependent manner, but do not influence the expressions of CD80 and CD86 in DCs; The therapeutic effect of sinomenine on rheumatoid arthritis may be related to its inhibition to secretion of IL-12 in DCs.

OBJECTIVE To investigate the effect of berberine on differentiation of rat bone marrow mesenchymal stem cells (MSCs) to adipocytes and its mechanism. METHODS Rat MSCs were isolated and cultured, adipocytic differentiation was induced with adipogenesis-inducing medium (AIM). Cells were assigned into 6 groups:normal control, AIM group, AIM+berberine 0.1, 0.3, 1 and 3 μmol·L-1 groups, respectively. Morphology characteristics of mesenchymal stem cells were observed under an inverted microscope and adipocyte levels were analyzed by oil O staining. Alkaline phosphatase (ALP) activity was detected using p-nitrophenyl phosphate as a substrate. The cell survival was determined by MTT assay. Expressions of peroxisome proliferator activated receptor γ (PPARγ), fatty acid binding protein (aP2) and CCAAT enhancer-binding protein α (C/EBPα) mRNA were detected by semiquantitative RT-PCR. RESULTS Compared with normal control group, MSCs adipogenic differentiation, PPARγ, aP2 and C/EBPα mRNA expression significantly increased in AIM group (P<0.01), ALP activity in AIM group significantly decreased (P<0.01). Compared with AIM group, berberine inhibited MSCs adipogenic differentiation (P<0.01) and berberine 0.1, 0.3, 1 and 3 μmol·L-1 increased ALP activity by 26%, 54%, 81% and 122%, respectively. Berberine 3 μmol·L-1 significantly downregulated PPARγ expression (0.91±0.10 vs 1.34±0.06) (P<0.01), aP2 (1.05±0.10 vs 1.53±0.09) (P<0.01) and C/EBPα mRNA (1.24±0.06 vs 1.54±0.09) (P<0.01). Berberine had no effect on proliferation of MSCs. CONCLUSION Berberine inhibits differentiation of MSCs into adipocytes, which might be closely related to the downregulation of PPARγ, aP2 and C/EBPα mRNA.

AIM To investigate the inhibitory effect of paeonol on hydrogen peroxide(H2O2)-induced apoptosis in PC12 cells. METHODS The injury model in PC12 cells was generated by H2O2 treatment. The cell viability was determined using methylthiazolyl tetrazolium reduction assay. Apoptotic cells and reactive oxygen species (ROS) were measured by flow cytometry. Lactate dehydrogenase (LDH) activity and malonyldialdehyde (MDA) content were measured by spectroscope respectively. RESULTS After PC12 cells were treated with H2O2 (100 μmol*L-1) for 10 h,its viability obviously decreased, and apoptotic cells, LDH release into the culture media, ROS and MDA contents in PC12 cells significantly increased. When the cells were pretreated with paeonol (12, 25 and 50 μmol*L-1)for 1 h prior to incubation with H2O2, its viability was greatly increased, and apoptotic cells, LDH release, ROS and MDA contents significantly decreased. CONCLUSION Paeonol protects PC12 cells from H2O2-induced apoptosis and this effect is probably achieved through its antioxidative action.

Objective To investigate the clinical guiding significance of lymphoscintigraphy with 99mTc-DX for radical lateral lymph node dissection with pelvic autonomic nerve preservation(PANP) in lower rectal cancer.Methods In 67 patients with pathologically proven lower rectal cancer,37MBq/0.8ml 99mTc-DX was injected into submucosa of the rectum through rectal endoscopy.At 0.5h,1h,2h,3h,4h,6h,12h,and 24h after the injection,the patients underwent pelvic and lower abdominal lymphoscintigraphy. The operative method was determined according to the imaging results,and the results of lymphoscintigrams were correlated with postoperative lateral node histologic examination.Results The scintigrams were postive in 37 cases,and all were unilateral.The positive patients underwent PANP.The image results were compared with histological lymph node examination in all patients operated upon for rectal cancer.Histologically,the cases in conformity were 26(70.3 %),false positive in 11 cases(29.7 %),and 2 of 30 false positive patients who demonstrated metastases to lateral node were operated by PANP,while the others underwent only PANP(type Ⅰ) without lateral lymph nodes dissection.The percentage of good sexual function of the two groups of cases after operation was 74.4 % and 71.4 % respentively.The percentages of mild dysuria and good function of urination were both 100 %.Conclusions Lymphoscintigraphy with 99mTC-DX has particular guiding significance in selection of PANP for lower rectal cancer.

Objective To investigate the evolution of pancreaticoduodenectomy and its significance in different time periods. Methods The clinical data of 165 patients from 1988 to 2008 in the department of general surgery, Zhangzhou Municipal Hospital with pancreaticoduodenectomy were retrospectively analyzed.Among 165 cases, simple pancreatoduodenectomy (PD) were performed in 138 cases, pylorus preserving pancreatoduodenectomy (PPPD) were performed in 14 cases, and extended pancreaticoduodenectomy were performed in 13 cases. The methods of pancreato-enteric reconstruction in pancreaticoduodenectomy included 68 cases with binding pancreaticojejunostomy (Peng's type Ⅰ ); 61 cases with pancreaticogastrostomy; 30 cases with traditional pancreaticojejunostomy; and 6 cases with duct-to-mucous pancreaticojejunostomy.Results From 1988 to 1998 (the first 10 years), 50 patients underwent pancreaticoduodenectomy, including 42 cases of PD and 8 cases of PPPD, and no case of extended pancreaticoduodenectomy. The mean amount of blood loss was (620 ± 180)ml, mean amount of blood transfusion was (530 ± 120)ml, the mean operation time was (6.5 ±3.5)h. Anastomotic fistula occurred in 7 cases, the incidence of anastomotic fistula was 14.0%(7/50); and 2 cases died during perioperative period with a mortality rate of 4.0% (2/50). From 1999 to 2008 (the latter 10 years), 115 patients underwent pancreaticoduodenectomy, including 96 cases of PD, 6 cases of PPPD, and 13 cases of extended PD. The mean amount of blood loss was (360 ± 110)ml, mean amount of blood transfusion was (400 ± 6 ) ml, the mean operation time was ( 3.0 ± 2.5 ) h, Anastomotic fistula occurred in 4 cases with an incidence of 3.5% ( 4/115 ); and one case during perioperative period with a mortality rate of 0.61%. The postoperative follow up time was ranging from 6 months to 5 years in 109 patients, the 1, 3,5 year survival rate was 87.2%, 54.1% and 39.5%. Conclusions In the latter 10 years, the amount of blood loss, the operation time, the mortality, and the incidence of pancreatic fistula have decreased significantly compared to the first 10 years.

Background and purpose:Cancer of unknown primary (CUP) represents approximately 5%~10%of malignant neoplasms. For CUP patients, identiifcation of tumor origin allows for more speciifc therapeutic regimens and improves outcomes.Methods:By retrieving the gene expression data from ArrayExpress and Gene Expression Omnibus data repositories, we established a comprehensive gene expression database of 5 800 tumor samples encom-passing 22 main tumor types. The support vector machine-recursive feature elimination algorithm was used for feature selection and classiifcation modelling. We further optimized the RNA isolation and real-time quantitative polymerase chain reaction (RTQ-PCR) methods for candidate gene expression proifling and applied the RTQ-PCR assays to a set of formalin-fixed, paraffin-embedded tumor samples.Results:Based on the pan-cancer transcriptome database, we identiifed a list of 96-tumor speciifc genes, including common tumor markers, such as cadherin 1 (CDH1), kallikrein-re-lated peptidase 3 (KLK3), and epidermal growth factor receptor (EGFR). Furthermore, we successfully translated the microarray-based gene expression signature to the RTQ-PCR assays, which allowed an overall success rate of 88.4% (95%CI: 83.2%-92.4%) in classifying 22 different tumor types of 206 formalin-fixed, paraffin-embedded samples. Conclusion:The 96-gene RTQ-PCR assay represents a useful tool for accurately identifying tumor origins. The assay uses RTQ-PCR and routine formalin-ifxed, paraffn-embedded samples, making it suitable for rapid clinical adoption.

Objective To investigate the pathogen distribution and risk factors of pulmonary infection after acute cervical spinal cord injury (ACSCI) in an attempt to offer reference for early antiinfection therapy.Methods The study comprised 223 cases who were admitted from October 2011 to October 2014.There were 149 males and 74 females,at (43.3 ± 13.5) years of age.Species of pathogens identified were gram-positive,gram-negative and mixed.Effects of age,gender,injury types and tracheotomy on pathogen distribution were analyzed.Results Gram-negative infection was found in 114 cases (51.1％),with tracheotomy accounting for 7.0％ of the cases and death accounting for 1.8％ of the cases,and the main causative pathogens were Klebsiella pneumonia,Escherichia coli,Pseudomonas aeruginosa and Acinetobacter baumannii.Gram-positive infection was found in 41 cases (18.4％),with tracheotomy accounting for 12.2％ of the cases and death accounting for 7.3％ of the cases,and the main causative pathogens were Staphylococcus aureus and Streptococcus pneumonia.Mixed infection was found in 68 cases (30.5％),with tracheotomy accounting for 22.1％ of the cases and death accounting for 13.2％ of the cases.Gender had no significant correlation with pathogen distribution.For the cases of complete spinal cord injury and tracheotomy,the ratio of mixed infection increased significantly (P ＜ 0.05).For the cases younger than 30 years,the pathogens were mainly gram-positive bacteria (P ＜ 0.05).Conclusions Main pathogens of pulmonary infection after ACSCI are gram-negative bacteria.The cases younger than 30 years are associated with higher risk of grampositive infection,while the cases with complete injury or tracheotomy are associated with higher risk of mixed infection.

Objective To analyze the effect of mesporous calcium silicate (m-CS)/calcium sulfate cement (CSC),m-CSC for short,in bone defect repair.Methods Setting time and compressive strength of the m-CSC (15 m-CSC as group Ⅰ and 30 m-CSC as group Ⅱ) were tested.CSC was used as the control.Cement samples were immersed in Tris-HCl solution,andin vitro degradation of the m-CSC was measured.Cell morphology and cell proliferation as well as differentiation on the samples were assessed.The cements were implanted into the traumatic femoral defects in rabbits,and the in vivo degradability and osteogenesis of the cements were investigated by histological evaluation after implantation for 4,8 and 12 weeks.Results Addition of m-CS into CSC prolonged the setting time (7.8 min in group Ⅰ and 10.5 min in group Ⅱ),obviously longer than 3.7 min in control group and did not have obvious effect on compressive strength of the cements.Weight loss of m-CSC solution was obviously lower (61.8 wt％ in group Ⅰ and50.3 wt％ in group Ⅱ),compared to70.4 wt％ in control group,pH value in group Ⅱ decreased from 7.40 to 7.26,while decreased from 7.40 to 6.86 in control group,m-CSC could promote cell proliferation and differentiation compared to CSC.At postoperative 12 weeks,histological sections showed massive new bony tissue (55.2％) in group Ⅱ,obviously higher than 25.6％ in control group.Conclusion m-CSC exhibits good biocompatibility,degradability and osteogenesis,and can promote bone regeneration in bone defect repair.

Objective To investigate the efficacy and feasibility of preimplantation genetic diagnosis (PGD) with human leukocyte antigen (HLA) matching for beta-thalassemia. Methods A total of 43 referred beta-thalassemia couples, with at least on child in need of hematopoietic stem cell transplantation (HSCT), underwent PGD for HLA matching at the First Affiliated Hospital of Sun Yat-sen University from 2010 to 2015. PGD cycles of these couples were retrospectively analyzed, and 15 infants born from PGD-HLA were followed up. Results A total of 84 oocyte retrieval cycles were performed, providing 14±7 oocytes per cycle. Fifty nine embryos biopsied cycles were done, including 24 cleavage stage and 35 blastocyst stage biopsy cycles. In cleavage stage, 259 embryos were biopsied, 93.4% (242/259) of them with conclusive molecular diagnosis, and the percentage of unaffected embryos (normo-homozygote and heterozygote) was 71.4%(185/259). The percentage of HLA matched unaffected embryos was 9.3%(24/259). In blastocyst stage, 306 embryos were biopsied, while 93.8% (287/306) of them were conclusive, and the percentage of unaffected embryos was 70.6% (216/306). The percentage of HLA matched unaffected embryos in blastocyst stage biopsy was 14.4%(44/306), which was higher than in cleavage stage biopsy (P<0.05). Forty three female carriers underwent 48 embryo transfer cycles including 3 fresh and 45 frozen-thawed embryo transfer cycles. Three fresh embryo transfer cycles were done after cleavage stage biopsy, resulted in a birth of healthy twins born at 36 weeks′gestation. All the embryos were frozen after blastocyst biopsied. Totally, 54 frozen-thawed embryos that were transferred in 45 frozen-thawed embryo transfer cycles included 25 embryo from cleavage stage biopsy and 29 embryo from blastocyst stage biopsy, and 42 of them were HLA matched. Clinical pregnancy rate and implantation rate per cycle in frozen-thawed embryo transfer were 38%(17/45) and 37%(20/54) respectively. A total of 15 infants were born, 2 were from a fresh embryo transfer cycle, and 13 were from frozen-thawed embryo transfer cycles. Results of prenatal diagnosis from delivered cases were matched to that of PGD. Four sick children have been cured by HSCT from these HLA matched born siblings. Conclusion PGD with HLA matching is an established method for conceiving a child who may donate hematopoietic stem cells to save an ill sibling.

OBJECTIVE: To determine the use of septal plication with Dor or Cooley procedure for post infarction anterior and anterior-septal aneurysm of the left ventricle. METHODS: A total of 23 patients with post infarction anterior and anterior-septal aneurysm of the left ventricle underwent septal plication and Dor or Cooley procedure along with coronary artery bypass grafting concomitantly. Data of NYHA grading, left ventricular end diastolic volume index (LVEDVI), left ventricular end systolic volume index (LVESVI) and left ventricular ejection fraction (LVEF) were recorded before the surgery, before discharge and 3 months after the surgery. RESULTS: Compared with the preoperative data, the NYHA grading before the discharge and 3 months after the surgery improved from 3.21 ± 0.62 to 1.72 ± 0.31 and 1.57 ± 0.23(P<0.05); LVEDVI decreased from (102.31 ± 18.71) mL/m² to (62.11 ± 6.21) mL/m² and (54.63 ± 4.54) mL/m² (P<0.05); LVESVI decreased from (69.32 ± 17.48) mL/m² to (30.23 ± 3.25)mL/m² and (28.34 ± 3.12) mL/m²; while LVEF increased from (32.92 ± 8.12)% to (48.78 ± 4.51)% and (50.52 ± 4.68)% (P<0.05), respectively. CONCLUSION Ventricular septal plication combined with Dor or Cooley procedure can remarkably improve the left heart function in patients with post infarction ventricular aneurysm.
Aged ; Coronary Artery Bypass ; methods ; Female ; Heart Aneurysm ; etiology ; surgery ; Heart Ventricles ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; complications ; Ventricular Function, Left ; physiology ; Ventricular Septum ; surgery