To compare the quality of cultivated species with w ild species of Desmodium styracifolium (Osbeck) Merr. TLC and UV were used to study their qualities. Results showed that there was no obvious di fference between them.

AIM:To investigate the inhibitory effect of aliskiren on the angiogenesis of human umbilical vein endothelial cells ( HUVECs) induced by lipopolysaccharide ( LPS) and to explore its possible mechanism.METHODS:HUVECs were cultured and randomly divided into blank group and renin group.The levels of tumor necrosis factor-α(TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in the culture supernatant were detected by ELISA.The protein levels of Toll-like receptor 4 ( TLR4) and ICAM-1 in the HUVECs were determined by Western blot.HUVECs were cul-tured and randomly divided into control group, LPS group, low-dose (1μmol/L) aliskiren group, middle-dose (10μmol/L) aliskiren group and high-dose (100 μmol/L) aliskiren group.The proliferation of HUVECs was detected by MTT and BrdU assays.The mobility of HUVECs was measured by Transwell assay.The formation of the vessels was judged by ob-serving the formation of the luminal structure by HUVECs in Matrigel.The levels of TNF-α, ICAM-1 and monocyte chemo-tactic protein 1 ( MCP-1) in the culture supernatant were measured by ELISA.The expression of renin, TLR4, matrix me-talloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) at mRNA and protein levels in the HUVECs was determined by RT-PCR and Western blot.RESULTS:Renin stimulated the expression of inflammatory factors and TLR4 in the HUVECs.Aliskiren inhibited the growth, migration and angiogenesis of HUVECs in a dose-dependent manner, de-creased the levels of TNF-α, ICAM-1 and MCP-1 and the expression of renin, MMP-2 and MMP-9, and inhibited TLR4 expression (P<0.05).CONCLUSION:Aliskiren inhibits LPS-induced angiogenesis of HUVECs, which may be related to the down-regulation of renin expression, the inhibition of TLR4-mediated inflammatory reaction, and the formation of MMP-9 and MMP-2.

Objective To prospectively evaluate the safety and therapeutic efficacy ofdalteparin in patients with high risk non-ST-elevation acute coronary syndromes (ACS) during percutaneous coronary intervention (PCI). Methods Atotal of 175 patients with high risk non-ST-elevation ACS were randomly assigned to 2 groups [dalteparin group and unfractionated heparin (UFH) group]. The patients in dalteparin group were given dalteparin at a dose of 5,000U subcutaneously soon after diagnosis and then an additional 60U/ kg intravenous bolus ofdalteparin before emergent PCI. Vascular access sheaths were removed immediately after PCI or coronary artery angiography; the patients in UFH group were given UFH intravenously at a dose of 25mgjust before PCI and an additional 65mg bolus was administered if angiographic findings showed that the patients were suitable for percutaneous transluminai coronary angioplasty (PTCA). Sheaths were removed at 4-6 hours after PCI; Results Eighty-three patients in dalteparin group underwent PCI while 82 patients in UFH group underwent PCI; anti-Xa activities of 52 patients in dalteparin group were measured. The average anti-Xa activity was (0.83±0.26) U/ml at 15 minutes after intravenous injection of dalteparin and anti-Xa>0.5U/ml was obtained in 96.1% of the patients; hematomas at puncture sites were significantly fewer in dalteparin group as compared with UFH group (2.3% vs 9. 2%, P < 0.05); none of the patients in 2 groups suffered major bleeding events. No death, acute arterial reocclusion or emergent revascularization events occurred at 30 days after PCI. Conclusions Our study demonstrated that early subcutaneous injection of dalteparin at a dose 5,000U after diagnosis and an additional 60U/kg intravenous bolus ofdalteparin before PCI is safe and efficacious for patients with high risk non-ST-elevation ACS undergoing emergent PCI

Objective To study the effect of the expansive open-door laminoplasty on the three-dimensional motion and stiffness of the cervical spine. Methods 55 cases after open-door laminoplasty of cervical spine due to myelopathy were follow-up for an average of 35.9 months. Cervical axial symptoms, neural functionality (JOA scoring system) and pre- and post-surgery dynamic cervical spine lateral X-ray films were evaluated. The Three-dimensional Motion and Stiffness of the Cervical Spine of Human Body Measuring Equipment was used in 12 post-operative open door laminoplasty cases and 10 pre-operative cases to measure the active and passive range of motion(ROM), load-displacement relationship, stiffness of cervical spine and torque caused by the isometric contraction of the extensors and flexors of the cervical spine. Results The average rate of improvement was 66. 2%. The excellent and good rate was 78.2%. The number of cases with distinct or severe cervical axial symptoms increased after the surgery ( P ＜ 0. O1 ) . The patients who have severe axial symptoms tend to have less curved cervical spine (P ＜ 0. 01 ) . There was no significant statistic difference between the severity of post-surgery cervical axial symptoms and JOA improvement level (P ＞ 0. 05). The active and passive ROM of extension, rotation and lateral bending of cervical spine were decreased after the surgery ( P ＜ 0. 05), and the main affection was on the middle and lower part of the cervical spine. The load-displacement figure of cervical spine can fit into an exponential equation T= b0eb1θ. In each direction, the lateral bending had the strongest stiffness, then the extension and flexion. The stiffness of rotation was the weakest. The stiffness of cervical spine of the post-surgery group was stronger than that of the contrast group. Conclusion The open-door laminoplasty of cervical spine damages the static mechanic balance on the sagittal plane and decreases the ROM and the flexibility of cervical spine. The curvature of the cervical spine is related to the severity of the axial symptoms and seems not to be related to the JOA score improvement.

Objective　To investigate the expression of endometrial leukaemia inhibitory factor（LIF） gene in patients with unexplained infertility. Methods　 By a quantitative reverse transcription-polymerase chain reaction (RT-PCR)，the expression of LIF gene on endometrium during mid-luteal phase was detected in 35 unexplained infertility (infertility group) cases and 20 infertile cases due to tubal obstruction or male factor (control group). Results　The level of LIF mRNA expression on endometrium during mid-luteal phase in infertility group was 0.448±0.239，significantly lower than those in the control group （1.093±0.761，P<0.01). Conclusions　Our findings suggested LIF might play an important role in the process of implantation. The decreased expression of LIF gene might be one of the major causes of unexplained infertility.

BACKGROUND: Beside direct trauma, a series of secondary pathological changes would occur in local injured spinal cord area during spinal cord injury. It has been reported that recombinant human erythropoietin (rhEPO)could inhibit cell apoptosis and inflammatory reaction, and possess neuroprotective role.OBJECTIVE: To explore the neuroprotective role of rhEPO in spinal cord injury by observing the nerve cell apoptosis and related cytokine expression in traumatic spinal cord.DESIGN: Randomized and controlled study.SETTING: Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, and Department of Pathology of Tongji Medical College, Central China Science and Technology University.PARTICIPANTS: The study was conduced at Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, Central China Science and Technology University and Department of Pathology of Tongji Medical College from September 2003 to May 2004. Thirty healthy female adult rats were randomly divided into 4 groups: ① Six rats in blank control group only subjected to spinal cord exposure without injury. ②Eight rats in injury group were subjected to spinal cord injury without medication. ③ Eight rats in medication A group were treated with rhEPO.④ Eight rats in medication B group were treated with thEPO and βaeacine sodium.METHODS: ① Animal model preparation: Spinal cord injury model was established on 24 rats by using improved Allen method. ② Administration: Rats in medication A group were treated with rhEPO in dosage of 300 U/(kg·d) at postoperative 1, 3, 5, 8, 11 days, while rats in medication B group were given additional β-aeacine sodium in dosage of 0.1 mg/kg,once a day for consecutive 11 days. Rats in blank control group and injury group were injected with the same volume physical saline from tail veins.③ Neurological function: The neurological function was scored by the same examiner at 1 day and 12 days after model establishment. Behavioral observation: Basing on improved Gale's neurological functional behavioral analysis, 0 score presented severer dysfunction and 6 scores represents normal. Slope test: The gradient was determined by the grasping capability of rat, which could reflect the recovery of neurological function. ④ Pathological examination: Rat was put to death at postoperative 12 days; traumatic spinal cord was made into slices for HE staining. ⑤ Expression of apoptosis cytokine bcl-2, bax and fas in nerve cells: Specimen was collected for IHC stain and brown colored positive cells was counted for calculating positive expressing rate. ⑥ Apoptosis nerve cells: In situ end-labeling techniques was used to calculate apoptosis index (the number of apoptosis nucleus/total number). All data were analyzed by using paired chi-test.MAIN OUTCOME MEASURES: ① Neurological functional behavioral score and results of slope test. ② Histological observation of traumatic spinal cord. ③ Expression of apoptosis nerve cell cytokines. ④ Examination of apoptostic nerve cells.RESULTS: ① Neurological functional behavioral score and results of slope test: There was no significant difference between 1 day and 12 days in blank control group, while the gradient in traumatic group was significantly larger in 12 days than in 1 day (P ＜ 0.05). In both therapeutic A and B groups, the behavioral scores and slope gradient were found significantly larger at 12 days than 1 day (P ＜ 0.05) and that of traumatic group (P ＜ 0.05). ② Histological observation of traumatic spinal cord: Traumatic spinal cord became slim with a majority of necrosis and glial cell hyperplasia in it; most of neurological tissues were found normal in medication A and B groups, with the neural structure of medication B group better than A group. ③Expression of nerve cell apoptosis cytokines: bax and fas positive cells in traumatic group were more than medication group A and B [traumatic group of (25.75±3.37)% and (41.37±2.83)% vs medication group A of (19.87±3.56) and (26.00±3.29)% vs medication group B of (12.00±2.97)and (17.50±2.20)％, P ＜ 0.05]; bcl-2 was significantly lower in medication group A and group B [(9.75±1.83)％, (14.63±2.83)％, (21.63±5.34)％,P ＜ 0.05]. ④ Examination of apoptotic nerve cells: Cell apeptosis index in traumatic group was significantly higher than medication group A and group B (50.75±5.39, 34.75±3.01, 24.00±3.46, P ＜ 0.05).CONCLUSION: Cell apoptosis is an important kind of neuronal death following spinal cord injury. rhEPO can inhibit nerve cell apoptosis and possess neuroprotective effect for traumatic spinal cord.

To modify the surface property of poly lactide-co-glycolide (PLGA) by biomimetic mineralization to construct a new kind of artificial bone. PLGA films and 3-diamensional (3-D) porous scaffolds hydrolyzed in alkaline solution were minerilized in SBF for 14 days. The morphology and composition of the mineral grown on PLGA were analyzed with SEM, FTIR and XRD. The porosity of the scaffolds was detected by using the liquid displacement method. The compressive strength of the scaffolds was detected by using a Shimadzu universal mechanic tester. An obvious mineral coating was detected on the surface of films and scaffolds. The main component of the mineral was carbonated hydroxyapatite (HA) similar to the major mineral component of bone tissues. The porosity of the un-mineralized and mineralized porous scaffolds was (84.86 +/- 8.52) % and (79.70 +/- 7.70) % respectively. The compressive strength was 0.784 +/- 0.156 N/mm2 in un-mineralized 3-D porous PLGA and 0.858 +/- 0.145 N/mm2 in mineralized 3-D porous PLGA. There were no significant differences between the mineralized and un-mineralized scaffolds (P > 0.05) in porosity and biomechanics. Biomimetic mineralization is a suitable method to construct artificial bone.
Biocompatible Materials ; Bone Substitutes ; Calcification, Physiologic ; Durapatite/metabolism ; Lactic Acid/*chemistry ; Polyglycolic Acid/*chemistry ; Polymers/*chemistry ; Porosity ; Tissue Engineering

ObjectiveTo observe the expression of apoptosis factor, the mechanism of neuronal apoptosis and profective effect of recombinant Human Erythropoietin (rHu EPO) after spinal cord injury (SCI) in SD rats.Methods30 SD rats were divided into four groups including control group, SCI group, treatment groups A and B.The animal SCI model was established with Allen's method. The changes of nerve functions of rats of 4 groups were observed before and after treating with rHu EPO. The expressions of apoptosis factors (Bcl-2,Bax,Fas) were tested with immunocytochemistry technique, and apoptosis neurones were labeled with TUNEL dyeing.ResultsThe grade of nerve function was improved distinctly in treatment groups, but group B was better than group A.The number of the positive cells for Fas and Bax in SCI group was more than that in treatment groups (P<0.05), and group A was more than group B. The number of Bcl 2 positive cells in the treatment groups was greater than that in SCI group (P<0.05).ConclusionApoptosis is a important death mode of neuron after SCI. rHu EPO can distinctly improve the comeback of the nerve function and protect the nerve tissue.

BACKGROUND: How to deal with bone defect is a big problem to surgeons. In recent years, the development in the technology of molecular biology and tissue engineering provides broad prospect for the clinical treatment of bone defect, which is one of the important study directions in department of orthopedics. The transforming growth factor-beta 1(TGF-β1),one of the important factors in bone formation, plays an important role in bone metabolism and recovery.OBJECTIVE: To observe the therapeutic effects of bone defects with osteoblasts transfected . By TGF-β1 combining with biomimetic biodegradable polymer scaffolds.DESIGN: Randomized controlled trial.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Siderophilin, trypsin, 3H-proline and sirius red, etc.MATERIALS: The experiment was completed in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from March to August in 2003. Twenty healthy adult Sprague-Dawley(SD) rats of SPF grade, weighing 200-250 g, were provided by the Experimental Animal Center of Tongji Medical College.METHODS: The osteoblasts transfected by TGF-β1 gene, combining with poly-DL-lactic acid scaffolds modified with poly-L-lysine, were transplanted into rat tibia defect. Radiographs and histological analysis were performed to evaluate the repair effects.MAIN OUTCOME MEASURES: The X-ray evaluation and histology observation were performed at the 4th and 8th weeks after the operation.RESULTS: Totally 20 SD rats were included in result analysis without one rat missing. ①In the experiment group, X-ray image indicated callus formation, while histology observation showed osteoid tissue and new bone formation, and osteoblasts attached to the surface of the materials after 4 weeks. Eight weeks later, the defect was essentially repaired, and the bone density of new bone was similar to that of the autogenous bone. ②In the control group, there was no formation of callus and osteoid tissue, and few osteoblasts attached to the surface of the materials, and a lot of lymphocytes infiltrated and blood capillary grew in the lacune of materials after 4 weeks. Eight weeks later, the imbedded materials were substituted mostly by fibrous tissue.CONCLUSION: The ideal repair effect of bone defect can be obtained through the combination of molecular biology with tissue engineering.

Objective To investigate the evolution of hepatitis C virus (HCV) quasispecies in persistent and self limited infection patients. Methods Peripheral blood HCV preserved for 10 years from 8 naive individuals once infected by HCV were analyzed. HCV hypervariable region 1 (HVR1) genes were cloned and sequenced. Entropy, genetic distances, nonsynonymous mutations per nonsynonymous site (K A ), synonymous mutations per synonymous site(K S ) were evaluated for evolutionary analysis. Results The entropy values and genetic distances of HCV quasispecies were lower in self limited infection indivi duals than that of persistent infection individuals. In 3 of the 4 persistent infection individuals there exist no difference between within group and between group genetic distances in HCV HVR1. The ratios of K A /K s were higher than 1 in 7 of these 8 individuals. Conclusions The complexity and diversity degree of HCV quasispecies may affect the outcome of HCV infection. Positive Darwinian selection may be the major cause for the change of HCV quasispecies distribution in hepatitis C infection natural history.