S. D rats were sensitized with PHA and were killed after two weeks. Peritoneal MC were collect-ed and separated using Arabic gum separating solution. Those MC were attacked by adding PHA andfol1owed by incubating at 37 ?C in water bath. After incubating for 5, 1O, l5, 2O, 30min and Ih,eachincubating medium was aspirated. The MC peroxidase was separated by disc electrophoresis and wasstained by ascrobic acid-benzidine stain. The relative peroxidase activity was calculated. Mean while,1O drops of 20 minute incubating medium were aspirated, fixed,embeded,sectioned,stainedand exam-ined under the electronic microscope routinely- 35 different kinds of 10 S. D. rat, s tissues were cut,embeded,sectioned,and stained with the toluidine blue and examined under light microscope for ob-serving the tissue distribution of MC. The results were as follows: (l )The peroxidase released fromthe PHA attacked MC was successfully demonstrated; (2)As comparing with the c0ntr0l group, theincreasing of the relative peroxidase activity in experirnental groups was not obvious at the intervals of5min and 10min incubating, but was obvious as the intervals of 15~3Omin incubating. No significantincreasing of the relative peroxidase activity was found at 60min incubating1 (3)When incubating for2Omin,the degranulation ratio of the experimental groups was 76%,while that of the control groupswas l6% only. There were two kinds of vacuoles in the degranulating MC plasrna, vacuoles withmembrane and without rnembrane. (4 ) The MC in skin, lung, respiratory tract, digestive tract, boneand thymus were prevalent. The order of MC number decreasing was as follows:urinary tract,repro-ductive tract, heart, liver, spleen, kidney. No MC was found in brain, spinal cord, hypophysis andtestls.

Objective To observe the counteraction of Ganoderma lucidum polysaccharide(GLP) on the inhibition of mice splenocyte proliferation and IL-1? and IL-2 mRNA expression induced by prostaglandin E2(PGE2).Methods Mixed lymphocyte culture reaction(MLR) method was used for the experiment.The lymphocyte proliferation was determined by MTT method,and the levels of IL-1? and IL-2 mRNA expression were evaluated by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results After co-culture with PGE2 for 48 hours,the splenocyte proliferation was inhibited to some extent,and the difference was significant when the concentration of PGE2 was over 10?mol/L(P

AIM:To study the relationship between apoptosis, proliferation and expression,mutation of related genes in breast cancer. METHODS: Methods of TUNEL, immunohistochemical S-P and PCR-SSCP were used respectively to study apoptotic index (AI), mitotic index(MI), expression of Bcl-2,p53,c-erbB-2,PCNA,Ki67,TopoⅡ and mutation of p53 in 54 cases of breast cancer. RESULTS: AI and MI were 9.40?3 78 and 5 96?2 36, respectively. There was a significant direct correlation between them( r= 0 46 P

[ ABSTRACT] AIM:To investigate the protective effect of 1, 3-dicyclopentyl-1, 2, 3, 6-tetrahydropyrimidine-4, 5-dicarboxylic acid diethyl ester (ZL-5015) on lethal endotoxin-challenged mice and to explore the underlying mechanism. METHODS:Mouse model of lethal endotoxin challenge and endotoxemia were established by intraperitoneal administration of lipopolysaccharide (LPS) at a dose of 70 mg/kg to the C57BL/6J mice.Mouse peritoneal macrophages stimulated with LPS (10 mg/L) were used as an in vitro inflammatory model.The levels of interleukin-1β( IL-1β) , interleukin-10 ( IL-10) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to evaluate the mRNA expression of the cytokines.RESULTS:Prophylactic treatment of the mice with ZL-5015 (100 and 200 mg/kg, ig) slightly increased the survival rate, extended the survival time, decreased the serum levels of IL-1βand TNF-α, and increased the serum level of IL-10 in the early stage of endotoxemia as compared with model group.The results of in vitro study demonstrated that treatment of the endotoxin-stimulated mouse peritoneal macrophages with ZL-5015 (10, 20 and 40μmol/L) inhibited the expression of IL-1βand TNF-αat both mRNA and protein levels but promoted the expression of IL-10 at both mRNA and protein levels.CONCLUSION: The tetrahydropyrimidine derivative ZL-5015 shows a moderate anti-endotoxin effect by increasing the survival rate and extending the survival time of the mice challenged by endotoxin, which may result from inhibition of the expression of pro-inflammatory cytokines such as IL-1βand TNF-α, and promotion of the expression of anti-inflammatory cytokine IL-10.

Basic knowledge of forensic science is theoretical and abstract,therefore,confir-matory experiment teaching is often adopted in the experiment teaching. International education has become an important topic in medical colleges and universities. Forensic medicine is a compulsory subject for medical international students. This article discussed the exploration and practice in teach-ing forensic medicine for international students at Wenzhou Medical University,which included faculty constructing and training,encouraging whole English teaching by young teachers,applying heuristic interactive teaching and case teaching,establishing strict attendance and appraisal system and con-ducting comprehensive evaluation on medical teaching quality and teaching management.

Objective To investigate the arsenic methylation level of people chronically exposed to different levels of arsenic in drinking water.Methods A cluster sampling method was used to select 874 cases that had drank different concentration arsenic-contaminated water from arsenic endemic area in Bayannaoer City.They were divided into four groups according to arsenic exposure level:control (≤ 10 μg/L),low (＞ 10-50 μg/L),medium (＞ 50-200 μg/L) and high groups (＞ 200 μg/L),146,155,224,349 cases,respectively.The content of arsenic in drinking water and the arsenic species in urine were analyzed by inductively coupled plasma mass spectrometry (ICPMSS) and the results were expressed as median.Results The inorganic arsenic (iAs),monomethylarsonic acid (MMA),dimethylarsinic acid (DMA),and total-arsenic (tAs) in urine of low,medium and high groups increased following increasing of arsenic exposure level (x2 =605.08,609.96,615.83,628.64,all P ＜ 0.017) and iAs％,MMA％ and MMA/DMA significantly increased following increasing of arsenic exposure level (x2 =112.30,56.60,86.47,all P ＜ 0.017).DMA％,PMI and SMI significantly decreased following increasing of arsenic exposure level (x2 =125.80,112.30,86.47,all P ＜ 0.017).In the four groups,iAs％ of female were 11.39％,12.28％,13.47％ and 17.58％,they were significantly lower than those of male's (15.52％,16.19％,17.45％,21.86％,Z =-4.22,-3.79,-4.60,-6.71,all P ＜ 0.05);and DMA％ were 76.95％,74.05％,72.76％,and 68.64％ in the four groups respectively,and the PMI of female were 0.89,0.88,0.87,and 0.82,both DMA％ and PMI were significantly higher than those of male in each group (71.17％,69.39％,67.36％,61.29％,0.84,0.84,0.83,0.78,Z =-4.00,-3.34,-5.50,-7.24,-4.22,-3.79,-4.60,-6.71,all P ＜ 0.05).In control group,arsenic metabolites levels of people were not significantly different in the three age groups (all P ＞ 0.05).Compared to the ≤30 age group,the MMA,DMA and tAs of 31-45 age group increased and DMA,DMA％,PMI of ≥46 age group increased while iAs％ decreased in high group (μg/L:72.71 vs 109.13,307.90 vs 419.50,505.59 vs 684.60,307.90 vs 418.26;64.31％ vs 68.45％,0.79 vs 0.83,20.71％ vs 17.35％,x2 =10.72,10.24,8.20,10.24,9.89,20.96,20.96,all P ＜ 0.017).Compared to the 31-45 age group,DMA％ and PMI of ≥46 age group increased while iAs％ decreased (64.91％ vs 68.45％,0.80 vs 0.83,20.14％ vs 17.35％,x2 =9.89,20.96,20.96,all P ＜ 0.017).Conclusion There is a significant dose response relationship between arsenic metabolites and arsenic exposure level,and arsenic methylation is related to gender and age.

Aim To investigate the effect of ligustrazine on pulmonary arterial pressure,thromboxane A2(TXA2) and prostacyclin(PGI2) in chronic hypoxic hypercapnic rats.Methods Thirty rats were randomly divided into control group(A), hypoxic hypercapnic group(B) and hypoxic hypercapnia+ ligustrazine (lig) group (C) .Results The mPAP in group B was significantly higher than that in group A(P0.05) ; The specific value of WA/TA(vessel wall area/total area) and SMC (the density of medial smooth muscle cells) were significantly higher in group B than that in group A(P0.05).Conclusion Ligustrazine can inhibit pulmonary hypertension and structural remodeling of pulmonary artery in chronic hypoxic hypercapnic rats by inhibiting synthesis of TXA2.

AIM: To investigate the effect of diltiazem on mean pulmonary arterial pressure(mPAP) and nitric oxide synthase(NOS) in arterioles in chronic hypoxic hypercapnic rats. METHODS: Twenty-four rats were randomly divided into three groups: control group(A),hypoxic hypercapnic group(B), hypoxic hypercapnia+ diltiazem group (C), constitutive endothelial NOS(ceNOS) were observed in arterioles of rats using the technique of immunohistochemistry,ceNOS mRNA were observed by the technique of in situ hybridization . RESULTS: (1)mPAP was significantly higher in rats of B group than that of A and C group( P 0 05),but mCAP was lower in rats of C group than that in B group.(2)Light microscopy showed WA/TA (vessel wall area/total area) was significantly lower in rats of C group than that of B group ( P

Objective To evaluate the efficacy and safety of imatinib in chronic myeloid leukemia (CML) patients and analyse the factors affecting the survival. Methods 135 CML patients receiving imatinib were evaluated for hematologic, cytogenetic, and molecular responses and adverse events. Results The median follow-up was 20 (range 3-67) months. The rate of cumulative complete hematological response (CHR), major cytogenetic response (MCyR), complete cytogenetic response( CCyR ) and complete molecular response (CMoR) in chronic phase CML patients were 97.9 %, 78.3 %, 72.2 % and 35.1%, respectively.These rates were significantly higher in chronic phase than in accelerated phase and blastic phase (P ＜0.001).The rate of CCyR in low-risk patients was significantly higher than high-risk patients (P =0.048). The estimated overall survival (OS) rate at 1, 3 and 5 year for chronic phase patients were (97.8±1.5) %, (95.2±2.4) % and (91.9±3.2) %, respectively. The estimated progression-free (PFS) survival rate at 1, 3 and 5 year were (92.6±2.7) %, (85.5±3.7) % and (81.3±4.3) %, respectively. The OS rate for accelerated phase patients at 6, 12 and 24 month were (93.8±6.1) %, (72.5±11.8) % and (64.5±12.9) %, the PFS rate were (92.3±7.4) %,(64.5±14.7) %, (53.7±15.7) %, respectively. The OS rate for blastic phase patients at 6, 12 and 19 month were (86.4±7.3) %, (45.4±11.4) %, (19.4±9.8) %, the PFS rate were (70.1±12.6) %, (37.6±15.6) % and (18.8±15.4) %, respectively. The OS and PFS of patients in chronic phase who achieved CCyR or CMoR were better than patients only achieved CHR (P ≤0.001). Multivariate analysis for survival of chronic phase patients indicated that imatinib resistance was the unfavourable factor for PFS (P =0.000, RR =46.744) and OS(P =0.007, RR =20.270). The non-hematological toxicity of imatinib was slight and tolerable, severe hematological toxicity was the major reason for dose reduction or drug discontinuation. Conclusion The efficacy of imatinib in chronic phase CML patients is significantly superior to which in accelerated phase and blastic phase; Achieving CCyR even CMoR is the most important thing for longer survival, iinatinib resistance is the major problem in the treatment with imatinib.

OBJECTIVETo optimize the experimental model of nitric oxide (NO) production in mouse peritoneal macrophages in response to lipopolysaccharides (LPS) stimulation.

METHODSMouse resident peritoneal macrophages were collected by lavaging the peritoneal cavity of mice with Hank's solution and stimulated with Pseudomonas aeruginosa LPS for NO production. NO concentration in the culture supernatants was measured with Griess Reagent. The influences of cell density, LPS concentration, LPS stimulation duration and culture medium volume on NO production were investigated. Finally, the feasibility of the model was confirmed with specific anti-inflammatory drugs.

RESULTSThe density of macrophages produced the most significant effect on NO production (P<0.001), and optimal results were obtained at the macrophage density of 6×10(6) cells/ml with a volume of 100 µl in each well in 96-well plate. At a LPS concentration below 1 µg/ml, NO production increased proportionally with the increment of LPS concentration (P<0.001), but the increment of NO production declined obviously at LPS concentrations beyond 1 µg/ml, and the peak NO production occurred at a LPS concentration of 10 µg/ml. NO production also increased significantly with the prolongation of LPS stimulation (P<0.05), and the increments were greater within 24-48 h than those in 48-72 h. NO content in the culture supernatant was associated with the medium volume, and the highest level occurred in a system volume of 100 µl. Aspirin (1 mmol/L), dexamethasone (10 µmol/L), and cyclosporin A (10 µmol/L) all significantly inhibited LPS-stimulated production of NO in mouse resident peritoneal macrophages (P<0.001).

CONCLUSIONSMacrophage density, LPS concentration, and the duration of LPS stimulation are the main factors affecting LPS-stimulated NO production in mouse resident peritoneal macrophages. The optimal results can be obtained with a macrophage density of 5×10(6) cells/ml (100 µl per well), LPS concentration of 10 µg/ml, LPS stimulation duration of 24 h or 48 h, and a culture medium volume of 100 to 200 µl.

Animals ; Cells, Cultured ; Female ; Lipopolysaccharides ; pharmacology ; Macrophages, Peritoneal ; drug effects ; secretion ; Male ; Mice ; Mice, Inbred Strains ; Nitric Oxide ; biosynthesis