We have carried out experiment design and comment,and students write the reports of experiment design in patho- physilolgy teaching from the aspects of basic process of experiment research and basic factors,principle and meaning of experi- ment design.By way of the teaching reform,the major position of students in studying is established,students' ability to study in- dependently and acquire knowledge actively are well cultivated,their comprehensive quality are enhanced and the teachers con- struction is also promoted.

The tandem expression vector can effectively increase the expression level and structure stability of the target peptides(protein)with small size of molecular,and avoid toxicity towards host cell from some toxic peptides product,therefore,this approach is widely applied in biotechnology.The four construction methods of tandem expression plasmid including asymmetric and complementary cohesive ends,directional adapter,isocaudarners,and tandem expression cassette were reviewed in terms of protocol,characteristic and applicable field.In addition,selection principles from various construction methods of tandem plasmid were reviewed in terms of efficiency,accuracy and product cleavage.

OBJECTIVETo evaluate the efficacy and progression-free survival of erlotinib after progression of disease to gefitinib in patients with advanced pulmonary adenocarcinoma who previously obtained a disease control with gefitinib.

METHODIn this retrospective study, 12 patients with advanced or metastatic pulmonary adenocarcinoma,who were previously obtained a partial response or a stable disease with gefitinib,were treated with erlotinib after gefitinib failure. Erlotinib efficiency, progression-free survival and overall survival were analyzed.

RESULTSNice (75%)achieved stable disease and three (25%) achieved progression disease with erlotinib treatment after gefitinib failure. No complete response or partial response was observed. The disease control rate was 75%. The median progression-free survival and overall survival of erlotinib were 180 days and 831 days.

CONCLUSIONErlotinib seems to be an optional treatment after gefitinib failure for advanced pulmonary adenocarcinoma patients,who previously responded to gefitinib.

Adenocarcinoma ; drug therapy ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Drug Tolerance ; Erlotinib Hydrochloride ; Feasibility Studies ; Female ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Quinazolines ; therapeutic use ; Retrospective Studies ; Treatment Outcome

Animals ; Blood Pressure ; Lymphatic Vessels ; physiopathology ; Male ; Norepinephrine ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; physiopathology

OBJECTIVETo explore surface roughness of bone cement and surround tissue on histological characteristic of induced membranes.

METHODSBone cements with smooth and rough surface were implanted in radius bone defect, intramuscular and subcutaneous sites of rabbits, and formed induced membranes. Membranes were obtained and stained (HE) 6 weeks later. Images of membrane tissue were obtained and analyzed with an automated image analysis system. Five histological parameters of membranes were measured with thickness,area,cell density,ECM density and microvessel density. Double factor variance analysis was used to evaluate the effect of the two factors on histological characteristics of induced membranes.

RESULTSMembranes can be induced by each kind of bone cement and at all the three tissue sites. In histological parameters of thickness,area and micro vessel,there were significant differences among the membranes induced at different tissue sites (P = 0.000, P = 0.000, P = 0.000); whereas, there were no significant differences in histological parameters of cell density and ECM density (P = 0.734, P = 0.638). In all five histological parameters of membranes, there were no significant differences between the membranes induced by bone cements with different surface roughness (P = 0.506, P = 0.185, P = 0.883, P = 0.093, P = 0.918).

CONCLUSIONSurround tissue rather than surface roughness of bone cements can affect the histological characteristics of induced membranes. The fibrocystic number, vascularity, mechanical tension and micro motion of the surround tissue may be closely correlated with the histological characteristics of induced membranes.

Animals ; Bone Cements ; Female ; Membranes ; cytology ; Rabbits ; Radius ; cytology ; Surface Properties ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective To explore the biological functions of retinoic acid-inducible gene-I(RIG-I) in vivo through phenotype analysis of RIG-I knockout mice. Methods The gene expression of RIG-Ⅰ in various tissues of mice was examined with Northern blotting and semi-quantitative RT-PCR.The phenotypes observed included body weight measurement,differential count of peripheral blood cells,metabolic parameters measurement and histopathologic examination. ResultsRIG-Ⅰ expressed in various tissues of mice with different levels.No gross developmental abnormalities and expected maturation arrest in granulocytic differentiation were observed in RIG-Ⅰ knockout mice.However,RIG-Ⅰ knockout mice exhibited an unexpected increase in the ratios of neutrophiles to lymphocytes in peripheral blood and increased susceptibility to bacteria infection. Conclusion RIG-Ⅰ may play an important role in immune regulation in mice.

Adult ; Aged ; Decompression, Surgical ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Nerve Block ; methods ; Spinal Nerve Roots ; Spinal Stenosis ; surgery

AIMTo observe the effect of mesenteric lymph duct ligation on free radical and inflammatory mediator of myocardium with severe hemorrhagic shock in rats at different period, and explore the effect of intestinal lymphatic pathway on myocardium injury pathogenesis in shock rats.

METHODS78 male Wistar rats were divided into the sham group, shock group and ligation group. The model of serious hemorrhagic shock was established in shock group, ligation group, and mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group after resuscitate. All rats were executed and taken out heart making for homogenate of 10 percent to determine the MDA, SOD, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6), myeloperoxidase (MPO), NO and NOS at after shock 90 min, after transfusion and resuscitate 0 h, 1 h, 3 h, 6 h, 12 h and 24 h etc. different times, and the expression of inducible nitric oxide synthase (iNOS) mRNA in myocardium was detected by RT-PCR.

RESULTSThe contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of shock group were rising after transfusion and resuscitate, and that was higher level at 3 h to 12 h, and that was significantly higher than sham group, the activity of SOD was significantly lower than sham group. The contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of ligation group were significantly lower than that of shock group at sameness points, and the SOD activity was higher.

CONCLUSIONThe mesenteric lymph duct ligation and blocking mesenteric lymph could reduce the PMN detaining, decrease the discharging of TNFa and IL-6, reduce the NO and expression of iNOS mRNA, and reduce the releasing of free radical and consuming of SOD.

Animals ; Free Radicals ; metabolism ; Inflammation Mediators ; metabolism ; Interleukin-6 ; metabolism ; Lymphatic Vessels ; metabolism ; Male ; Mesentery ; metabolism ; Myocardium ; metabolism ; pathology ; Neutrophils ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; pathology ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

The aim of the present study was to investigate whether protein kinase C (PKC) was involved in the effect of mesenteric lymph duct ligation or mesenteric lymph drainage on vascular calcium sensitivity in hemorrhagic shock rats. Male Wistar rats were randomly divided into Sham, Shock (hemorrhagic shock), Shock+Ligation (mesenteric lymph duct ligation plus shock) and Shock+Drainage (mesenteric lymph drainage plus shock) groups. After being in shock (hypotension 40 mmHg) for 3 h, the tissue of superior mesenteric artery (SMA) was taken out for detecting the PKC expression and phospho-PKC (p-PKC) activity, and the vascular rings of SMA were prepared and used to measure the response to gradient calcium concentration for assaying the calcium sensitivity, the parameters of which including tension, maximum tension (E(max)) and negative logarithm of EC(50), called the pD(2). Other vascular rings from Shock+Ligation and Shock+Drainage groups were incubated with PKC regulator PMA or Staurosporine before the measurement of calcium sensitivity. The results showed that, PKC expression, p-PKC activity and calcium sensitivity of SMA in Shock group was significantly lower than that of Sham group, whereas the above-mentioned indexes were significantly elevated in Shock+Ligation and Shock+Drainage groups compared with those in Shock group. PKC agonist PMA enhanced the contractile activity of vascular rings to gradient calcium ions, and increased E(max) of SMA in Shock+Ligation and Shock+Drainage groups. On the contrary, PKC inhibitor Staurosporine significantly decreased the response to gradient calcium ions and E(max) of SMA in Shock+Ligation and Shock+Drainage groups. These results suggest that PKC plays a role in the improvement of vascular calcium sensitivity by blockade of mesenteric lymph return in hemorrhagic shock rats.
Animals ; Calcium ; pharmacology ; Drainage ; Ligation ; Lymph ; physiology ; Lymphatic Vessels ; physiology ; Male ; Mesenteric Artery, Superior ; drug effects ; physiology ; Mesentery ; Muscle, Smooth, Vascular ; drug effects ; metabolism ; Protein Kinase C ; metabolism ; physiology ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology ; Vasoconstriction ; drug effects ; physiology

The aim of the present study was to investigate the changes of lymphatic contraction after hemorrhagic shock in vitro and the underlying role of nitric oxide (NO). Rat thoracic duct segments were isolated at 0, 0.5, 1, 2 and 3 h after hemorrhagic shock. Using Pressure Myograph System, we determined contraction frequency (CF), end systolic diameter (ESD), end diastolic diameter (EDD) and passive diameter (PD) of isolated rat lymphatics under different transmural pressures (1, 3, 5, 7 and 9 cmH(2)O), then calculated contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) of lymphatics. The results showed that in several transmural pressures, lymphatic CF, TI and FPF were significantly higher in shock 0 h and shock 0.5 h groups than those in control group (sham operation group). With the development of shock, lymphatic CF, TI and FPF decreased significantly in shock 2 h and shock 3 h groups compared with those in control group. We further discovered the role of NO in the changes of lymphatic contraction after hemorrhagic shock. Under 3 cmH(2)O transmural pressure, the changes of lymphatic contraction in shock 0.5 h and shock 2 h groups were analyzed following the incubation with several NO-related drugs alone or in combination. And the results showed that NO donor L-Arg reduced CF, TI and FPF in shock 0.5 h group to the control levels, while soluble guanylate cyclase inhibitor ODQ suppressed the effect of L-Arg. Moreover, NOS inhibitor L-NAME elevated the CF, TI and FPF of 2 h shock lymphatics to the control levels, while phosphodiesterase inhibitor aminophylline (AP) suppressed the effect of L-NAME. These results suggest that the lymphatic contractile activity exhibits a biphasic change during hemorrhagic shock, increasing in early phase and declining in later stage. And NO plays a major regulating role in the biphasic change of lymphatic contraction in hemorrhagic shock rats via cGMP pathway.
Animals ; Cyclic GMP ; metabolism ; In Vitro Techniques ; Male ; Muscle Contraction ; Muscle, Smooth ; physiopathology ; Nitric Oxide ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology ; Thoracic Duct ; physiopathology ; Time Factors