With that general outlook, the relationship between Y in and Y ang is left and right relationship. Con-scious spirit and primordial spirit are a couple of important concepts in traditional Chinese medicine (TCM). Firstly, Conscious spirit is the function of the highest part of brain while primordial spirit is the function of the lower-level part of the brain. The function of highest part of brain is represented by the frontal lobe and cerebral cortex. In conclusion, from the development and evolution point of view, conscious spirit is corresponded with post-natal aspects while primordial spirit is corresponded with prenatal aspects. Secondly, conscious spirit is Y ang while primordial spirit is Y in . Motion is Y ang while stillness is Y in . Daytime is Y ang while nighttime is Y in .Conscious spirit is mainly appeared in reaction and daytime. Primordial spirit is mainly appeared in the rest and nighttime. Hot is Y ang while cold is Y in. Primordial spirit is mainly appeared in the sleep and rest when the en-ergy metabolism degrades. Therefore, from the viewpoint of conscious spirit and primordial spirit, the relationship between Y in and Y ang is up and down relationship.

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.

BACKGROUND:It has been reported that treatment with granulocyte-colony stimulating factor (G-CSF) increases the abundance of circulating CD34+ cells in rats. Data from the study, more important, suggested that mobilized by G-CSF may enhance rapid reendothelization and reduce neointimal formation after vascular injury.OBJECTIVE: To evaluate whether BM-derived CD34+ cells could enhance rapid reendothelization and reduce neointimal formation after balloon-injured carotid artery in an intact rat model.DESIGN: Randomized control animal study.SETTING: Nanjing First Hospital Affiliated to Nanjing Medical University.MATERIALS: The experiment was carried out in Nanjing First Hospital from December 2005 to April 2006. A total of 40 male Sprague-Dawley rats, weighing 200-250 g and of SPF grade, were purchased from National Rodent Laboratory Animal Resources, Shanghai Branch. The recombinant human G-CSF was purchased from Qilu Pharmaceutical. The 2F Fogarty arterial embolectomy catheters were purchased from Edwards Lifesciences. Anti-human CD34 and anti-human CD45 were purchased from Multi Sciences.METHODS: SD rats were divided randomly into treated group (n =20) and control group (n =20). Subcutaneous injection of recombinant human G-CSF (100 μg/kg/day) once daily for 8 days for treated group. Control group as treated with subcutaneous injection of saline. Five days after initiation of G-CSF treatment or saline, the rats were anesthetized by intraperitoneal injection with ketamine. The left common carotid artery was exposed through a midline incision of the ventral side of the neck. A 2F Fogarty arterial embolectomy catheter was inserted through the external carotid artery,inflated with 200 μL air, and passed 3 times along the length of the segment, which was defined proximally by the carotid bifurcation and distally by the edge of the omohyoid muscle. After removal of the catheter, the proximal ligature of the external carotid artery was tied off. ① An average of 1 mL venous blood per rat was collected for enumeration of the white blood wells (WBCs) and CD34+ cells before and 5 days after initiating G-CSF or saline treatment. ② Ten rats in each group were killed with overdose ketamine at 14 and 28 days after balloon injury and left common carotid arteries were harvested. The luminal surface of carotid arteries (n =5, each group) was exposed to calculate the reendothelialized area, which was manually traced with software (Image ProPlus). Reendothelialized area = non-stained with Evans blue area/the total area of balloon-injuried. The cross sections of carotid arteries (n =5, each group) were stained with hematoxylin and eosin (HE) and calculated intima-to-media area ratio (I/M) with software (Image ProPlus) to assess the extent of neointimal thickening. ③ To evaluate the extent of reendothelialization of arteries injury, sections were stained with CD31 and vWF by immunohistochemistry analysis.MAIN OUTCOME MEASURES: ① The number of WBCs and CD34+ cells; ② the extent of reendothelialization of arteries injury; ③ the extent of neointimal hyperplasia (I/M); ④ CD31 + and vWF+ endothelial cells.RESULTS: A total of 40 rats were involved in the final analysis. ① The number of WBCs and CD34+ cells: After 5 days of treatment, the number of WBCs in the treated rats increased more than 2.7-fold compared with control group [(27.60±2.45) ×109 L-1, (10.11±1.81) ×109 L-1, P ＜ 0.01], CD34+ cells increased more than 12.2-fold compared with control group (38.31×107 L-1, 3.14×107 L-1, P ＜ 0.01). ② The extent of reendothelialization: At 14 and 28 days after balloon injury,carotid artery of reendothelialization in the treated group were (68.3±8.3)％ and (97.6±4.1)％, superior than the control group (33.8±6.3)％ and (76.1±5.2)％ (P ＜ 0.01). ③ The extent of neointimal hyperplasia: At 14 and 28 days after balloon injury, the neointima-media (I/M) ratios in the treated rats were 0.39±0.11 and 0.45±0.09, less than the control group 0.87±0.15,1.26±0.16 (P ＜ 0.01). A highly significant inhibition of neointimal hyperplasia was observed in the treated group. ④ CD31+ and vWF+ endothelial cells: At 28 days after injury, sections from G-CSF treated group showed almost complete and continuous monolayer of CD31 and vWF positive cells.In contrast, a patchy and interrupted CD31 and vWF positive cells were found lining the lumen of control group.CONCLUSION:Treatment with G-CSF significantly increases the number of CD34+ cells and accelerates the rate of reendothelialization of injured vessels, leading to marked inhibition of neointimal formation after vascular injury in rats.

In the study,nematode-trapping fungus-Arthrobotrys oligospora was firstly cultivated in Sabouraud dextrose broth medium containing 0.05％ of agar,then transferred to the corn meal agar medium.A.oligospora conidiospores was eluted from the media in different time and lyophilized after being counted,then the resuscitation of lyophilized spores was also observed,in oder to evaluate their nematicidal dosage and nematode-trapping efficacy in vitro.The results of the study were as follows:by observing the germination rate,growth rate and nematode-trapping rate of lyophilized spores from A.oligospora.The maximum germination rate of lyophilized A.oligospora conidiospores was 79.5％ on the 4 th day after inoculation,and the average growth rate was 3.4 mm/d;the maximum nematode-trapping rate was 95.8％ on the 7 th day after larvae were added on the media,and the average nematode-trapping rate was 74.0％.Compared with the control groups,the differences were both no significant (P＞0.1)in average growth and nematode-trapping rate.The results show that the freeze-dried preparation materials was accessible and simple,with good resuscitation.After further optimization it will display the prospect of industrialization application.

Objective To observe the effects of melatonin on synaptic plasticity impaired by spinal cord injury in rats. Methods A total of 54 female Sprague-Dawley rats were divided into sham group (n=18), control group (n=18) and melatonin group (n=18). Spinal cord inju-ry model was established with modified Allen's method at T10 (10 g from 25 mm height). The number of neurons and the expression of the Nissl body were detected with immunofluorescence and Nissl staining. The expression of neurofilament-200 (NF-200), brain-derived neuro-trophic factors (BDNF), Synapsin I and growth-associated protein-43 (GAP-43) was detected with Western blotting. Results Seven days af-ter injury, the number of motoneurons, the expression of Nissl body in motoneurons, and the expression of BDNF, Synapsin I and GAP-43 decreased in the control group compared with those in the sham group, and they increased in the melatonin group compared with those in the control group. Conclusion Melatonin can repair the impaired synaptic plasticity, which might promote the functional recovery after spi-nal cord injury.

Objective To investigate the influence of matrix metalloproteinases-9 (MMP-9) on permeability of blood-spinal cord barrier (BSCB) after spinal cord injury (SCI). Methods 68 male C57BL/6 mice were randomly divided into sham group, 2 days group (S2), 7 days group (S7) and 14 days group (S14) after SCI with 17 mice in each group. All the groups received a moderate impacted spinal cord injury except the sham group. Evan's Blue (EB) was administered intraperitoneally to detect the permeability of BSCB. Occludin was analyzed by immunofluorescence, the expressions of occludin and MMP-9 were detected by Western blotting. Results After SCI, BMS score significantly reduced, compared with S2 group, S14 group showed a significant increase (P<0.01). The permeability of BSCB was seriously damaged after SCI. Compared with S2 group, S14 group showed a notable down-regulation in the permeability of injured micro-vessels (P<0.05). The expression of occludin was down-regulated and the expression of MMP-9 was up-regulated 7 days after SCI (P<0.05). Compared with S2 group, S14 group showed a significant up-regulation of occludin and a remarkable down-regulation of MMP-9 (P<0.05). Conclusion After SCI, MMP-9 might mediate the expression of occludin to influence the BSCB permeability.

AIM:To study the effect of gossypol on the cognitive function of type 2 diabetic rats,and to explore its mechanism. METHODS:Thirty male Sprague-Dawley rats were divided into three groups randomly:normal group,type 2 diabetic group and gossypol treated group. After fed with high-fat diet for 4 weeks,the later two groups were injected with streptozotocin intraperitoneally to establish type 2 diabetic rat model. The animals in gossypol treated group were given gossypol at dosage of 15 mg/kg once per day for 4 weeks by gavage. Since 5th week,the times of gavages were changed into once per week at the same dosage and lasted to 12th week. Learning and memory abilities of rats were assayed with Morris water maze test. The concentration of blood glucose was measured by biochemical method. The levels of serum corticosterone and insulin were detected by enzyme-linked immunosorbent assay and radioimmunoassay,respectively. The protein expressions of 11?-HSD1 and GR in cerebral cortex and hippocampus were determined by Western blotting. The morphological changes of cerebral cortex and hippocampus were observed under light microscope and transmission electronic microscope,respectively. RESULTS:Compared to normal group,the karyopyknosis,dilation of golgiosome and mitochondria swelling of neuron from cerebral cortex and hippocampus were prominent in diabetic group. The concentrations of blood glucose,serum corticosterone and insulin increased significantly (P

Objective To study the differences between the animal model of pulmonary injury/ fibrosis induced by using paraquat and that induced by using bleomycin in mice in order to establish an ideal mouse pulmonary fibrosis model.Methods Thirty healthy and 8 ～ 10 weeks old male C57BL/6J (C57) mice were randomly (random number) divided into paraquat group (n =10),bleomycin group (n =10),and control group (n =10).Paraquat ( 10 mg/kg) was given to mice intraperitoneally once every three days for 5 times in paraquat group.Bleomycin was injected into trachea of mice in a dose of 3 mg /kg in bleomycin group.The mice were sacrificed 7 days,14 days and 21 days after administration of drug.The general physical condition,body weight and pulmonary pathological changes were observed.Data were analyzed with SPSS13.0 statistical package.The comparison was made between two groups with mann -whitney U- test.Results Both agents could induce pulmonary injury and fibrosis.After comparison of survival rate,body weight,pulmonary histopathological change and rate of successful modelling,the repeated low - dose of paraquat injected intraperitoneally was proved to be a method of more simple and effective with high success rate of modeling in comparison with the conventional technique of intratracheal injection of bleomycin.Conclusions By the comparison between two methods of establishing pulmonary injury and fibrosis models in mice,the method of repeated low - dose intraperitoneal injection of paraquat is superior over the bleomycin - induced method in respect of higher rate of successful modelling.

Objective According to the requirements of the College of American Pathologists (CAP),to validate the laboratory developed test-JAK2 V617F mutation.Methods This is a clinical laboratory diagnosis study.Fifty-seven peripheral blood and bone marrow samples were collected from the BCR/ABL-negative myeloproliferative neoplasms (MPN) patients in Huashan Hospital from Apr 2010 to Dec 2010.JAK2 V617F mutation was detected by fluorescence PCR combined with melting curve analysis.The accuracy,precision,analytical sensitivity,and anti-interference ability of the assay were validated.The Kappa test was used to evaluate accuracy.Coincidence rate was used to evaluate the repeatability.Paired t test was used to evaluate analytical sensitivity.Results There was a close agreement between the reference method(sequencing) and the test method (fluorescence melting curve analysis) (Kappa =0.89).The coincidence rate was 100％.The results of the assay was not affected by lipoprotein (＜ 24 mmol/L) or bilirubin (＜450 μmol/L).The detection limit of the assay was 5％.Conclusion JAK2 V617F mutation assay by fluorescence PCR combined with melting curve analysis could be applied to the clinical laboratory.

Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.