Objective: To observe effects of Yuzhang Decoction on in vitro proliferation of bone marrow CD_(34) cells in the patient of aplastic anemia (CAA) and its mechanism. Methods: To adopt monoclone antibody-immune magnetic beads separation system (MACS) to separate and purify CD_(34) cells in bone marrow of the patient of CAA, and make cell culture, applying ~3H-TdR incorporation to detect effect of Yuzhang Decoction on proliferation of CD_(34) cells of bone marrow. Results: CD_(34) cells of bone marrow showed concentration-dependent proliferation in the Yuzhang Decoction group with significantly differences (P

Co2EDTA-DMAP.HCl.

The biologically active constituents in Gynostemma pentaphyllum are dammarane-type glycosides,called gypenosides.They are believed to be the highest contents of this herb,easy to obtain,and mainly active in anti-tumor,controlling the blood glucose,lipid-lowering,cardiovascular protection,etc.The saponins may change into sub-glucoside after hydrolysis,for the intemal acetal glucoside structure is vulnerable to acid,alkali,and enzyme degradation.Through searching the literatures in recent years,this paper summarized the chemical constituents in the hydrolyzed products of gypenosides,for providing references to discover novel and more active lead compounds.

Objective:To construct an eukaryotic expression vector of enhanced green fluorescence protein(EGFP) gene driven by telomerase catalytic subunit(hTERT) gene promoter and observe the specific expression of EGFP in lung cancer cell lines.Methods:The 1100bp promoter fragment was obtained by enzyme digestion from a recombinant plasmid of pGL3-hTERTp containing the hTERT promoter.The hTERT promoter was then subcloned into the upstream of the report gene EGFP of pEGFP-1 without promoter.The expression vector pEGFP-hTERTp was successfully constructed.The vector pEGFP-N1 containing cytomegalovirus(CMV) promoter was used as a positive control.The vector pEGFP-1 without promoter was used as a negative control.The vectors were transfected into human lung cancer cell lines 95D,NCI-H446,A2,A549,LTEP-a-2,YTMLC and normal MRC-5 through lipofectamine respectively.EGFP expression was detected under the fluorescence microscope.Results:pEGFP-hTERTp was confirmed by enzyme digestion with correct result.That the EGFP expression was detected in all of eight lung cancer cells transfected with pEGFP-hTERTp,but not in MRC cells.By contrast,high intensity EGFP expression was observed in both lung tumor cells and normal cells,which were transfected with pEGFP-N1.Conclusion:The EGFP controlled by hTERT promoter can be expressed specifically in lung cancer cell lines.hTERT promoter may be used as an excellent regulation element in tumor-targeting gene therapy.

BACKGROUND:As a traditional treatment for multilevel cervical myelopathy,nterior long-segmental decompression has the shortcomings of great operative trauma,high difficulty,low fusion rate,etc.,which can affect the postoperative efficacy.OBJ ECTIVE:To evaluate the clinical effects of three different anterior surgeries on multilevel cervical myelopathy.DESIGN:A comparative observation.SETTING:Department of Orthopaedics,Changzheog Hospital,the Second Military Medical University of Chinese PLA.PARTICIPANTS:Thirty-six patients with multilevel cervical myelopathy of 3 consecutive segments,who were surgically treated,were selected from the Department of Orthopaedics,Changzheng Hospital,the Second Military Medical University of Chinese PLA from June 1999 to June 2003,including 25 males and 11 females,35-62 years of age,the disease course ranged from 3 to 26 months. According to the clinical manifestations and imaging esults,they were diagnosed as multilevel cervical myelopathy,and they were not suffering from consecutive ossification of posterior longitudinal ligament and ossification of ligamenta flava. Informed contents were obtained from all the patients and their relatives.METHODS:All the patients were grafted with utologous bone. Autologous ilium or cancellous bone excluding vertebral body was filled into titan net or Cage,which were made of titan and characterized by high intensity,tolerance to decay,good biocompatibility,etc. According to the operative manner,the patients were divided into 3 groups:① two-level corpectomy with fusion group(long-segmental decompression group,n =11):There were 4 cases grafted with long-titan net,and 7 cases grafted with autologous iliac bone. Sub-total two-level corpectomy with fusion was performed. ②segmental decompression group(n =16):including 12 cases of titan net+cage graft,4 cases of autologous bone+cage graft. One-level decompression and sub-total single corpectomy with fusion were performed. ③three-level decompression group(n =9):Only discectomy without corpectomy was performed. After complete decompression,3cages were used to fill artificial bone or grafted with autologous bone.MAIN OUTCOME MEASURES:Cervical anteroposterior and lateral radiographies,flexion and extension radiograph were reexamined within 1 week and at 3,6 and 12 months postoperatively. The neurological function was assessed using the Japanese Orthopaedic Association(JOA) scoring method preoperatively and 3 months postoperatively. The total score was 17 points,the higher the score,the better the neurological function. The duration of operation,perioperative bleeding amount,length of stay,cost of hospitalization,graft fusion at 3 months postoperatively,improved JOA score at 3 months postoperatively were recorded in the three groups. The occurrence of postoperative complications was observed by means of return visit.RESULTS:All the 36 patients with multilevel cervical myelopathy were involved in the analysis of results. The mean duration of operation,mean perioperative bleeding amount and mean length of stay in the segmental-decompression group and three-level decompression group were obviously fewer or shorter than those in the long-segmental decompression group(P ＜ 0.05),and the average cost of hospitalization was obviously higher than that in the long-segmental decompression group(P ＜ 0.05). The postoperative improved JOA score and graft fusion rate were close among the groups(P ＞ 0.05).CONCLUSION:Segmental anterior cervical decompression is a recommendable technique for multilevel cervical myelopathy by comprehensively considering the fusion rate,recovery of neurological function,duration of operation,perioperative bleeding and length of stay.

Objective To investigate the functional and ultrastructural changes of cerebral microvascular endothelial cells at early stage following traumatic brain injury (TBI) in rats. Methods The rat models with closed brain injury were established with the improved Marmarous method. The expressions of thrombomodulin (TM) and von Willebrand factor (vWF) were determined by immunohistochemical techniques (5 rats per group) and real-time quantitative RT-PCR (5 rats per group) respectively at 1, 4, 8, 12, 24 hours and at days 3 and 7 after injury. Results TM and vWF started expression at 4 hours, reached peak at 24 hours and recovered to normal at day 7 after TBI. The expression levels of TM and vWF at different time points in sham control group showed statistical difference compared with damage group (P < 0.05). Conclusion The activation of the cerebral microvascular endothelial cells at early stage after TBI is the main mechanism of early secondary brain injury.

Objective To assess the results of surgical intervention on patients with medullary thyroid carcinoma(MTC), and determine the value of measuring plasma calcitonin concentration postoperatively. Methods The diagnosis and treatment of 14 patients with MTC from January 1992 to December 1998 were analysed retrospectively. Results The diagnosis of MTC in the 14 patients was confirmed by pathology. Of them, 64.3% of patients had lymph node metastases. According to AJCC staging system, 1 patient was in stage Ⅰ, 7 in stage Ⅱ, 5 in stage Ⅲ and 1 in stage Ⅳ. Of nine patients measured plasma calcitoinin after initial operation, 4 had persisted hypercalcitoninemia. In the 4 patients, MTC in residual thyroid and enarged lymph node were comfirmed by B mode ultrasounography. After re operation, the calcitonin level returned to normal in 3 cases, but one remained in higher level. Postoperative follow up ranged from 2 to 8 years, 2 patients died of the disease. Twelve patients still lived, 6 of them survived more than 5 years. Conclusions The clinical stage of MTC at the time of diagnosis is an important prognostic factor. An aggressive surgical approach at the initial operation is essential to achieve a curative effect in patient with MTC. Measuring plasma calcitonin postoperatively helps to detect residuled MTC or recurrent MTC.

Objective To assess the clinical manifestation, management and outcome of abdominal aortic aneurysm (AAA) in young adult Methods Retrospective study was made on young AAA patients (

A CdTe/ZnSe quantum-dot submicrobead ( QBs ) , which exhibited fluorescence intensity approximately 2800-fold stronger than that of single quantum dots, was conjugated with the anti-histidine rich protein( HRP )-Ⅱ mAbs using N-( 3-( Dimethylamino ) propyl )-N'-ethylcarbodiimide hydrochloride ( EDC ) method as fluorescence probe. The goat anti-HRP-Ⅱ polyclonal antibodies and donkey anti-mouse polyclonal antibodies were sprayed onto the nitrocellulose membrane as test line and control line, respectively. The resultant fluorescence probes were introduced to the immunochromatographic strip for the quantitative determination of Plasmodium falciparum. For determination of Plasmodium falciparum in serum, the QBs based immunochromatographic strips exhibited a good dynamic linear range from 5 . 8 Parasite/μL to 8010 Parasite/μL with a limit of detection of 5. 8 Parasite/μL. The detection time of the proposed QBs based immunochromatographic strips for each sample was only 15 min. Moreover, the recovery rates of the intra-and inter-assay ranged from 93. 0% to 111. 9%, and 98. 3% to 115. 1% respectively, while the relative standard deviations ( RSDs) of intra-and inter-assay were below 5%.

BACKGROUND:The effects and molecular mechanism of simvastatin on the proliferation and differentiation of osteoblasts remain unclear. Especial y, we do not know much about the effects of connexin 43.
OBJECTIVE:To evaluate the effects of simvastatin on the proliferation and differentiation of osteoblasts and the regulatory effect of simvastatin on the expression of osteogenic genes and connexin 43.
METHODS:Newborn Sprague-Dawley rats were chosen and the cranium digestion method was used to culture osteoblasts. The different concentrations of simvastatin (0.062 5, 0.125, 0.25, 0.5 and 1.0μmol/L) were used to deal with osteoblasts. The proliferative effect of simvastatin on osteoblasts was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The effect of simvastatin on osteoblast differentiation was measured with alkaline phosphatase activities. The mRNA and protein expression of osteogenic genes and connexin 43 were measured by real time quantitative RT-PCR and western blot assay.
RESULTS AND CONCLUSION:There were no significant differences in absorbance values of simvastatin groups at 3 days (P>0.05). However, at 4 and 5 days, absorbance values were lower in the simvastatin groups than those in the control group (P<0.05). Compared with the control group, alkaline phosphatase activities of osteoblasts were greater in the simvastatin groups (P<0.05). Moreover, the effects of 0.25μmol/L simvastatin on alkaline phosphatase activities of osteoblasts were most significant. Osteocalcin, alkaline phosphatase activities, type I col agen and connexin 43 mRNA and protein expressions were increased after treatment with 0.25μmol/L simvastatin (P<0.05). These results indicated that simvastatin may inhibit the proliferation and improve the differentiation of osteoblasts by upregulating the mRNA and protein expression of osteogenic genes and connexin 43. These data may provide the new intervention target for osteoporosis treated with statins.