Objective To construct baculovirus expression system for the human plasminogen kringle 5(hK5) gene,and detect its expression in Spodoptera frugiperda 21(sf21). Methods Baculovirus Bacmid-rhK5 was prepared with bac-to-bac baculovirus expression system,and sf21 cells were infected.The expression of rhK5 in sf21 cells was determined by Western blot.The activity of purified protein was detected by vascular endothelial cell inhibition test. Results The pFastBac HTB-rhK5 was successfully constructed,and sf21 cells were transfected with the constructed vector.The output of rhK5 obtained was 90 ug/L,and the protein possessed the inhibitory activity of endothelial cell proliferation.The median effective dose(ED50) was 4 ug/mL. Conclusion The rhK5 baculovirus expression vector is highly expressed in sf21 cells,which lays a foundation for the the expression of rhK5 protein with biologic activities.

Objective To evaluate the gastroesophageal reflux symptom in patients who underwent continuous ambulatory peritoneal dialysis (CAPD) and the efficacy of proton pump inhibitor (PPI) in treating gastroesophageal reflux. Methods Fifty-eight CAPD patients with good clinical and complete dialyzed eondition,who was admitted to the hospital between Jan. 2008 and July 2008, were inquired about their gastroesophageal reflux symptoms using reflux disease questionnaire (RDQ). The patients who had RDQ≥6 and <12 were received esomeprazole 20 mg daily, while those with RDQ≥12 were received esomeparzole 20 mg twice daily. RDQ score was reevaluated 4 weeks after treatment.Results The common symptom was regurgitation (64.70%), followed by acid reflux (52.9 %), non-cardic chest pain (47.1. %) and heart burn (17. 6%). After 4-week treatment, the RDQ was significantly decreased (P< 0. 05). But there was no difference in outcome of treatment between patients with RDQ≥ 12 and RDQ< 12 (P=0. 059). Conclusion The gastroesophageal reflux symptom in CAPD patients can be relieved by PPI administration, but a larger clinical trial is needed to evaluate the course and efficacy of treatment.

This paper explored factors influencing initiatives of clinical teachers in millitary medical university for training clinical undergraduates including work pressure after being transferring to civilian,work pressure,policy orientation,economic pressure,student factor and psychological factor,etc. Meanwhile,this paper explored the incentive mechanism of clinical teachers in millitary medical university for training clinical undergraduates including training and evaluating clinical teach-ing teachers, innovating evaluation criterion and creating sustainable teaching atmosphere (building examples and establishing various incentive systems). All measures taken above was mean to inspire the initiatives of clinical teaching teachers and to ensure the quality of clinical teaching.

Objective To investigate the localization and the morphological changes of manchette during mouse spermiogenesis.Methods Immunofluorescence staining with FITC and costaining with DAPI were used to demonstrate the cellular localization of the manchette at different stages during mouse spermiogenesis.The structural changes of the manchette were observed during the maturing of the spermatid.Results Immunofluorescence staining showed that manchette existed exactly around the nuclei of the spermatids.Manchette began to form,when the shape of the nucleus changed from spherical to slightly elongated.While the nucleus of the spermatids condensed and elongated at later stages,manchette moved gradually to the caudal position of the spermatids.At last,the manchette diminished as the spermatids became mature.During mouse spermiogenesis,manchette underwent a transition from a cap-like to a tubular configuration.ConclusionThe formation and diminishment of the manchette is in step with the condensation and elongation of the nucleus of the spermatid.Both the structural and positional changes of the manchette coincide with the changes of the nucleus.These results imply that manchette might play an important role in mouse spermiogenesis.

Objective To investigate the protective effects of mild hypothermia on cells in the CA_1 region of the hippocampus in gerbils following global ischemia-reperfusion injury(IRI),and to explore their mechanism. Methods IRI models were established in 75 gerbils.Any changes in TUNEL positive cells and the expression of Bax and Cytochrome C were then observed in normothermic(N) ,hypothermic(H)and sham(S) groups through immu- nohistochemistry methods.Results In the H group(as compared with the N group)apoptotic cells in the CA_1 sub- field of the hippocampus significantly decreased.The expression of Bax and Cyt C at 3 h,6 h and 1 d were de- creased,and the expression fastigium was delayed.Conclusion Mild hypothermia can moderate and delay cell ap- optosis,and its mechanism might be related with reducing and delaying the expression of Bax and Cyt C released by mitochondria.

Over the last decade, chronic critically ill (CCI) has emerged as an epidemic in intensive care unit (ICU) survivors worldwide. Advances in ICU technology and implementation of care bundles has significantly decreased early deaths of critically ill patients, and have allowed them to survive previously lethal multiple organ failure (MOF). However, more and more survivors leave persistent low grade organ dysfunctions, depend on continues organ support, need to stay in ICU, and become CCI patients. These patients experience a persistent immune dysregulation with persistent inflammation, immunosuppression, and catabolic syndrome. Therefore, malnutrition is an important feature of patients with CCI, and nutritional support is a crucial part of their treatment. The main strategies of nutritional support are as follows: providing sufficient calories and proteins with appropriate anabolic agents to promote anabolic metabolism, using immunomodulators to improve immune suppression and inflammatory responses, and supplementing micronutrients to enhance metabolic support. In this review, the nutritional assessment, calorie assessment, protein assessment and other nutrient supplementation (such as β blocker, testosterone and oxandrolone, immunonutrition, vitamins) of CCI patients were reviewed, so as to provide reference for the treatment of CCI.

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

OBJECTIVETo explore the effect of Danggui Yinzi (DY) on delayed allergy in model mice with qi-blood deficiency syndrome (QBDS).

METHODSQBDS model was established in 48 Kuming mice of SPF grade by using reserpine and acetophenone hydrazine. Forty of them were then randomly divided into the model group, the loratadine group, the high dose DY group, the middle dose DY group, and the low dose DY group, 8 in each group. Another 8 in line with the same standard were recruited as a blank group. Mice in high, middle, and low dose DY groups were administered with DY concentrated solution at 60, 30, 15 g/kg by gastrogavage. Mice in the loratadine group were administered with loratadine solution at 1.66 mg/kg by gastrogavage. Equal volume of normal saline was administered to mice in the model group and the blank group by gastrogavage. All medication was given once per day for 1 successive week. Except those in the blank group, the rest mice were evenly smeared with 1% DNCB solution on the abdomen. Five days after skin allergy, 1% DNCB solution was smeared to right ear of all mice to stimulate allergic reaction. Mice in the blank group were smeared in the same way without allergenic reaction. The auricle swelling and the inhibition ratio were determined at 24 h after attack. Blood was collected from orbit and serum IgE level detected using double-antibody sandwich ELISA.

RESULTSCompared with the blank group, auricle swelling obviously increased and serum IgE level was obviously elevated in the model group (P < 0.01). Compared with the model group, auricle swelling obviously decreased and serum IgE level was obviously reduced in the 3 dose DY groups (P < 0.05, P < 0.01). Meanwhile, the auricle swelling degree was superior in high and middle dose DY groups to that in the loratadine group (P < 0.05). The inhibition ratio of auricle swelling was sequenced from high to low as 67.3% in the high dose DY group, 56.0% in the middle dose DY group, 48.1% in the low dose DY group, 47.3% in the loratadine group.

CONCLUSIONSDY could inhibit auricle swelling and lower serum IgE level. It also could inhibit delayed allergic reaction in model mice with QBDS to some extent.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Edema ; drug therapy ; Hypersensitivity, Delayed ; drug therapy ; Immunoglobulin E ; blood ; Loratadine ; pharmacology ; Mice ; Qi ; Random Allocation

Objective To investigate the role of GABAB1 receptors in spinal dorsal horn neurons in the development of diabetic neuropathic pain (DNP). Methods Sixty pathogen free male SD rats aged 4 weeks weighing 150-170 g were randomly divided into 2 groups ( n = 30 each): control group and DNP group. Diabetes mellitus was induced by single intraperitoneal injection of streptozotocin 50 mg/kg. Blood glucose levels and paw withdrawal threshold to mechanical stimuli were measured at 3, 5 and 7 weeks (T1, T2, T3 ) after IP STZ/NS ( n = 10 each). The animals were sacrificed after PWL measurement. The lumbar segment of the spinal cord was removed for determination of the expression of GABAB1 receptors by immuno-histochemistry, RT-PCR and Western blot analysis. Results The blood glucose levels were significantly higher while the PWT was significantly lower at T1,T2 and T3 in group DNP than in control group. The expression of GABAB1 receptor mRNA and protein in spinal dorsal horn was significantly lower at T2 and T3 in DNP group than in control group. Conclusion The expression of GABAb1 receptors is down-regulated in spinal dorsal horn neurons in rats with DNP.

Objective To evaluate the role of M3 muscarinic acetylcholine receptor (M3 mAChR) in the release of glycinergic neurotransmitter by using oxotremorine-M (Oxo-M: a nonselective mAChR agonist) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP: a highly selective M3mAChR antagonist). Methods Twenty male 3-4 weeks old SD rats weighing 160-180 g after successful intrathecal catheterization were randomized into 2 groups (n = 10 each): normal saline group (group NS) and pertussis toxin (group PTX).Pertussis toxin 1.5 μg/10 μl was injected IT in group PTX, while in group NS normal saline 10 μl was injected IT instead. The animals were killed at day 7 after injection. The spinal cords were removed and sliced and placed in artificial CSF. Glycinergic spontaneous inhibitory postsynaptic currents (sIPSCs) were measured in spinal lamina Ⅱneurons using whole-cell voltage-clamp technique. Five minutes after sealing, Oxo-M (final concentration 3 μ mol/L) was added. Oxo-M was then completely washed out 3 min later and 4-DAMP (final concentration 25 nmol/L) was added after 5 min of stabilization. In the presence of 4-DAMP, Oxo-M (final concentration 3 μmol/L) was added again 3 min later. sIPSCs were recorded before addition of Oxo-M (T1), 3 min after addition of Oxo-M (T2), 3 min after addition of 4-DAMP (T3), 3 min after the second addition of Oxo-M (T4). Results Compared with the baseline value at T1 , Oxo-M significantly increased the frequency of glycinergic sIPSCs at T2without changing the amplitude at T2-4 in both groups. The frequency of sIPSCs was significantly lower at T4 than at T2 in both groups (P ＜ 0.05 or 0.01). There was no significant difference in both frequency and amplitude of glycinergic sIPSCs between the two groups. Conclusion M3 mAChR plays a predominant role in the release of glycinergic transmitter in the spinal lamina Ⅱ neurons in rats.