[Objective]To study the effect of T cell groups of CD4+,CD8+,CD4+/CD8+,IL-4,IFN-? in the hypoimmunitymice induced by hydrocortisone to discuss its function of regulating the immune function and CD4+TH cell’s inferior change of group,have offered the experiment basis for clinical practice of TAF.[Method] FACS measured the T lymphocyte phenotype;the level of cytokine IFN-r and IL-4 was detected by ELISA.[Result] TAF can raise CD4+T lymphocyte of hypoimmunitymice,suppress CD8+ T lymphocyte expression,raise CD4+/CD8+ ratio;And can make the level of serum IFN-r obviously enhanced in hypoimmunitymice,and the level of IL-4 was very low.[Conclusion] TAF can rebuild immune function of hypoimmunitymice,promote organism Th0 to split up to Th1,maintain the advantage state of TH1,and strengthen the function of cellular mediated immunoresponse.

In this study gloxylic acid-induced fluorescent histochemical method was usedto reveal the innervation of adrenergic nerves,acetycholinesterase (AChE) methodto show the cholinergic nerves,and the immunohistochemical method to display thevasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) immunoreactivenerves.This observation on the vas deferens showed that the VIP-immunoreactivenerves had the same distribution as the cholinergic nerves,and mainly existed inlamina propria,while NPY-immunoreactive nerves had the same distribution asadrenergic nerves and mainly existed in the muscular layer.One group of animalswas treated by intraperitoneal injections of 6-OHDA on the lst,2nd,6th and 7thday,and killed on the 8th day.Following treatment with 6-OHDA,significantevanescence of most adrenergic nerves was observed.NPY-immunoreactive nervesreduced remarkably and the extent of the decrease was similiar to that of adrenergicnerves,whereas no change was observed on the VIP and cholinergic nerves.Theimmunoelectronic microscopy (PAP pre-embedding method) demonstrated that VIPimmunoreactive complex deposited in the varicosity containing small clear vesicles(diameter of 40-55nm).Some varicosities also contained small number of largegranular vesicles with the diameter of 100-144mm.NPY immunoreactive complexdeposited in the large granular vesicles,and occasionally can be observed in thesmall granular vesicles,40-55nm in diameter.The results from this study suggestthat VIP probably co-exist with ACh and NPY with noradrenalin in rat vasdeferens.

Objective To study the effect of hypoxia on the expression of interleukin 6 (IL 6) in cocultured pulmonary arterial smooth muscle cells (PASMC) Methods Rat PASMC were cocultured with rat lung microvascular endothelial cells, and randomly divided into 5 groups: normal group (N), 2 h hypoxia (H2), 6 h hypoxia (H6), 12 h hypoxia (H12), and 24 h hypoxia (H24) The expression level of IL 6 mRNA in PASMC was detected with RT PCR, and the activity of IL 6 in the supernatant with radioimmunoassay Results The expression level of IL 6 mRNA increased in PASMC in H2, reached the highest in H6, and decreased in H12, but still higher than that of N The changes of the IL 6 activity in the supernatant as well as IL 6 mRNA expression in the cells were in a time dependent manner Conclusion Hypoxia can enhance the expression of IL 6 in cocultured PASMC And it may activate others genes which regulate the hypoxic response of PASMC and signal transduction

Objective To investigate the distribution modes of NOS nerves and interstitial cells of Cajal (ICCs) in myenteric plexus of human fetal intestine. Methods Small intestine samples from 12 fetuses were collected for the preparation of whole mount preparations and cryo sections examined by immunocytochemistry for c kit receptor tyrosine kinase and histochemistry for NADPH diaphorase. Results ICCs associated with the myenteric plexus in the fetal small intestine were observed in the shape of spindle or ovoid with two to three slender processes forming an independent and complete cellular network. While NOS positive nerves, which constitute the main neuronal component of ganglia, connecting strands, and nerve fibers, were found within the circular muscle layer and myenteric plexus of the small intestine and the latter was especially rich of this kind of nerves. Most of these positive neurons were Dogiel I type neurons and they often gathered in cluster in the ganglia. Each ganglion contained several to dozens of NOS positive neurons. Although no colocalization of ICCs and NOS positive nerves was found by double staining of whole mount preparations and cryo sections, they were closely distributed. Conclusion Our results indicate that NO released by myenteric plexus, an inhibitory neurotransmitter, may possess the function of regulating ICCs and smooth muscle in late stage of the fetus.

Objective To investigate the correlation between meteorological factors and the incidence of acute cardiovascular disease.Methods The number of calls for ambulance in case of cardiovascular disease between 2006 to 2010 was obtained from Beijing 120 emergency center and the data of meteorological factor during for the same period from Beijing Meteorological professional were collected,and then the impact of the weather factors of different seasons of weather factors on the incidence of cardiovascular disease was ere analyzed by time-stratified case-crossover design.Results The average daily incidence cases of cardiovascular disease in four seasons average daily were 5.61,4.55,5.05 and 6.03 respectively.The association between temperature Temperature and the incidence of acute cardiovascular disease was were non-linear relationship changes in spring and autumn seasons.Atmosphere Air pressure and the incidence were negatively correlated in the winter.The relative humidity of airand the incidence were related in the spring,summer,autumn.Wind and the incidence were related in the spring,summer,but negative correlation in the autumn,winter.Conclusions The incidences of acute cardiovascular disease are higher more in winter and spring than in summer and autumn.The incidences of cardiovascular disease are related with temperature,relative humidity,wind speed and other meteorological factors.,The influence of meteorological factors on the incidence of the disease delays to occur after changes in weather.

By using the histochemical method, the electron microscopy and biochemical measurement, this study deals with the changes of the innervation of the uterus in tats induced by pregnancy. The results revealed that the adrenergic and cholinergic innervation of the uterus were reduced in advanced pregnancy. The content of noradrenaline(NE), expressed as ng/per gram wet weight was 194.79?2.85 in the control group and 78.56?0.48 in the late pregnancy group (P

Fibroblast growth factor -21 (FGF-21) is a recently discovered metabolic regulation factor, regulating glucose and lipid metabolism and increasing insulin sensitivity. FGF-21 is expected to be a potential anti-diabetic drug. Expression of FGF-21 as inclusion bodies has advantages for high yield and purity, but the bioactivity of the protein is almost totally lost after denature and renature. That is why FGF-21 is currently expressed in soluble form. As a result, the yield is considerably low. In this study, we used SUMO vector to express SUMO-human FGF-21 (SUMO-hFGF-21) in form of inclusion body. We optimized the culture conditions to increase the yield of the bioactive human fibroblast growth factor-21. We applied the hollow fiber membrane filtration column to enrich the bacteria, wash, denature and renature inclusion bodies. After affinity and gel filtration chromatography, we examined the hypoglycemic activity of FGF-21 by the glucose uptake assay in HepG2 cells. We also detected the blood glucose concentration of type 2 diabetic db/db model mice after short or long-term treatment. The results show that the yield of ihFGF-21 was 4 times higher than that of shFGF-21. The yield was 20 mg/L for ihFGF-21 vs. 6 mg/L for shFGF-21. The purity of ihFGF-21 was above 95%, while shFGF-21 was 90%. Compared with the traditional method of extracting inclusion bodies, the production cycle was about three times shortened by application of hollow fiber membrane filtration column technology, but the bioactivity did not significantly differ. This method provides an efficient and cost-effective strategy to the pilot and industrial production of hFGF-21.
Animals ; Bacteria ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Disease Models, Animal ; Fibroblast Growth Factors ; biosynthesis ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; Humans ; Hypoglycemic Agents ; isolation & purification ; Inclusion Bodies ; metabolism ; Mice ; Recombinant Fusion Proteins ; biosynthesis ; Small Ubiquitin-Related Modifier Proteins ; biosynthesis

Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.

Objective To investigate the effect of the eye-acupuncture therapy on serum TNF-α levels in rats with cerebral ischemia-reperfusion injury and the related mechanism. Methods Healthy SD rats were randomly divided into four groups: normal group, sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group according to body weight. Sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group were divided into the 3h group, the 24h group and the 72h group, a total of 10 groups (n=8). To establish the rat model of cerebral ischemia-reperfusion by suture method in ischemia-reperfusion model group and eye-acupuncture group. Eye-acupuncture was separately started immediately after reperfusion and at 30 min before sampling in the 3h eye-acupuncture group, besides, eye-acupuncture was separately taken every 12h in the 24h eye-acupuncture group and in the 72h eye-acupuncture group. The rats in normal group were not treated, those in sham-operated control group were inserted the fishing thread 1cm, and the others were identical with those in ischemia-reperfusion model group. At 3h, 24h, 72h after reperfusion, the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks in ischemia-reperfusion model group and eye-acupuncture group. The method of ELISA was taken to detect the change of serum TNF-α levels in rats after the eye-acupuncture therapy. Results Compared with the ischemia-reperfusion model group, the neurologic impairment score of the eye-acupuncture group decreased obviously; The levels of serum TNF-αin ischemia-reperfusion model group at 3 h, 24 h, 72 h respectively were(76.803±18.325)pg/ml、(85.511±13.334)pg/ml、(86.831±9.232)pg/ml. The level of serum TNF-α in normal group was(24.304±6.511)pg/ml. The level of serum TNF-α in Sham-operated control group at 3 h, 24 h, 72 h respectively were(24.928±3.792)pg/ml,(27.533±5.362)pg/ml,(29.366±5.874)pg/ml. Ischemia-reperfusion model group compare with normal group and sham-operated control group, the difference were significant(P＜0.01). The levels of serum TNF-α in rats after the eye-acupuncture therapy at 3 h, 24 h, 72 h respectively were(40.185±3.335)pg/ml, (48.523±7.687)pg/ml, (51.611± 6.403)pg/ml. Compared with ischemia-reperfusion model group, the levels of serum TNF-α were strongly reduced(P＜0.01). Conclusion The eye-acupuncture therapy could play a role in improving cerebral ischemia-reperfusion injury evidently and the mechanism was related to its reducing serum TNF-α levels.

Fibroblast growth factor-21 (FGF-21) is an important metabolism regulator, however, whether FGF-21 has effects on cardiovascular remains unclear. In this study, H2O2-induced injury in H9c2 cells was used as a cell model, the anti-apoptosis potential and mechanism of FGF-21 against oxidative injury were evaluated by MTT assay, flow cytometry assay and real-time PCR. The results showed that FGF-21 could increase the cell survival of H2O2-induced injury in H9c2 cells and prevent H9c2 cells from oxidative stress-induced apoptosis. Furthermore, FGF-21 can elevate SOD activity and regulate Bcl-2/Bax expression in H9c2 cells. The results suggest that FGF-21 have protective effect against the H2O2-induced apoptosis in H9c2 cells.