Objective To study general law of typing according to syndrome differentiation of Chronic Obstructive Pulmonary Disease (COPD). Method Based on large sample prospective Clinical Epidemiology investigation, symptoms, conditions of tongue, pulse according to TCM and pulmonary function, quality of life in 616 patients of COPD were collected. By means of Frequencies procedure, factor analysis, cluster analysis and analysis of variance in SPSS11.5, analyze the investigated data and lay difficient types of syndrome of COPD according to TCM. Meanwhile, explored distinct distribution and the relation between types of syndrome and pulmonary function, quality of life in patient5 with COPD. Result There were six types according to syndrome differentiation of COPD, deficiency of lung-qi, pathogenic wind-cold attacking lung, deficiency of pulmonosplenic both vital energy and yin, yin-deficiency of the lung and kidney, stagnation of phlegm and pathogenic fluid-reteneion in lung following splenic asthenia and stagnation of liver-qi, blood stasis-phlegm following deficiency of heart-yang and kidney-yang. There were multiple types of syndrome in COPD patients, the relation was detected between each type of syndrome and lung function, quality of life. Conclusion To draw assistance from clinical epidemiology and mathematical statistics method, the initial standard of differentiation of symptoms and signs to COPD was layed. The regularity of types of syndrome to COPD was detected. The result could be used in diagnosis of COPD, and the method could be communicated in the standardization research of syndrome differentiation.

OBJECTIVE: To study the general law of typing of bronchiectasis according to syndrome differentiation. METHODS: We collected the symptoms, conditions of tongue and pulse in patients of bronchiectasis, using frequencies procedure, discriminant analysis and K-means cluster analysis in SPSS statistical software as research medium. RESULTS: Five hundred and sixty three patients with bronchiectasis were studied. It suggested that accumulation of phlegm-heat in the lungs (45.65%), liver fire attacking the lungs (24.51%), asthenia of pulmonosplenic qi (22.38%), asthenia of both qi and yin (7.46%) were the main types. CONCLUSION: Clinical epidemiology provided scientific basis for further studying of the typing of bronchiectasis according to syndrome differentiation. Building up differentiation of syndromes through differentiation and analysis of main symptoms can be used in clinical diagnosis.

BACKGROUND: Spinal cord can regenerate after injury in certain microenvironment. Olfactory ensheathing cells (OECs)have the characteristics of astrocytes and Schwann cells and can accelerate the spinal cord axonal regeneration.OBJECTIVE: To make injured thoracic cord rat models and observe the effect of OECs on injured spinal cord axonal regeneration.DESIGN: Observational experiment.SETTING: Second Affiliated Hospital of Sun Yat-Sen University.MATERIALS: The experiment was performed at the Second Affiliated Hospital of Sun Yat-Sen University from January 2001 to November 2002.Totally 20 adult SD male rats with the body mass of (380±20) g were provided by Experimental Animal Center of Sun Yat-Sen University (number of institution license SYXK2004-0020). There were DMEM culture solution with low glucose (L-DMEM, GibcoBRL), fetal calf serum (FCS) (Hyclone), myelin basic protein (MBP) (Sigma) and nerve growth factor receptor antibody (Sigma). They were divided into cell transplantation group and control group by the method of random digits table with 10 in each group.METHODS: The adult SD rats were anaesthetized and decapitated to obtain the whole olfactory bulb and then isolate olfactory nerve with a sterile operation. Thoracic cord injury models were established by modified Allen method. 10μL OECs suspension (2.5×1010 L-1) was injected into injured spinal cord of the cell transplantation group, whereas DMEM/F12 (1:1) culture solution of the same dose was injected in the control group. The influence of OECs on spinal cord axonal regeneration was observed by histological and immunohistochemical method 6 weeks after transplantation.MAIN OUTCOME MEASURES: ①OECs were identified by nerve growth factor receptor antibody staining. ②Repair of myelin sheath was observed by MBP staining. ③Nerve axonal regeneration was observed by argentaffin staining. RESULTS: Two rats in the cell transplantation group and 3 rats in the control group died, so totally 15 rats were involved in the result analysis. ①In the cell transplantation group,injured spinal meninges were integrated,but spinal cord became thin as compared with the normal spinal cord. By hematoxylin-eosin (HE) staining, multipolar cells appeared in injured region and the cells were fused excessively with myeloid tissues. It proved that survival OECs were integrated well within the host. New axons were clustered in bundles and infiltrated by small round lymphocytes. By argentaffin staining, regenerated axons grew in tissues of injured region, which mostly accompanied with fascicular-arranged multipolar cells. In the control group, spinal cord became thin markedly and spinal meninges were integrated. No new axon appeared in the injured spinal cord by HE staining. No regenerated axon appeared by argentaffin staining, either. ②In the cell transplantation group, most multipolar cells were clustered in bundles. A mass of positive granules of nerve growth factor receptor antibody appeared in cytoplasm, which further verified that OECs still survived and integrated well within the host 6 weeks after transplantation. Linear MBP positive fibers appeared in the injured region by MBP staining,which indicated that myelin-like substance appeared and drew closely in both ends of injured region. At the same time,MBP positive substance also appeared in the multipolar cells, which illustrated that transplanted OECs could induce the occurrence of myelin-like substance. No regenerated axon was found in the control group.CONCLUSION: Delayed transplantation of OECs can survive and induce the occurrence of myelin-like substance in injured spinal cord of adult rats.

The paper was aimed to review the modeling methods of spleen-yang deficiency, which can be classified into imitating traditional Chinese medicine (TCM) etiology methods and modern medicine methods. At present, methods of modeling were not generally accepted and the main presented problems were as follows. Modern medicine methods did not conformed to the rules of TCM etiology and pathogenesis. The evaluation of yang deficiency models was not accurate. Some modeling methods can lead to other diseases at the same time. The names of spleen deficiency model were not unified. It was difficult to distinguish the modeling methods of spleen-yang deficiency and spleen-qi deficiency, and etc. Model evaluation of spleen-yang deficiency was various. However, the domestic standard evaluation system had not been formed. The main model evaluation problems were as follows. Macro symptoms of the model lacked of objective and quantitative evaluation. It lacked of evaluation indexes of pulse and tongue with TCM characteristics. The disproof of prescription selection was not unified. Similar syndromes had not been ruled out. The microscopic indexes were fragmented, and etc. This paper reviewed on animal modeling methods of spleen-yang deficiency.

Objective To observe the effects of cryopreserved olfactory ensheathing cells(OECs )transplantation on spinal cord axonal regeneration and spinal func tional improvement.Methods Twenty-four rats were divided into experimental a nd control groups,with each group of 12rats.The spinal cord in-jury was es tablished by transecting the spinal cord with microsurgery scissors.OECs were p urified from SD rat olfactory bulb and cultured in DMEM and cryopreserved(-1 20℃)for 2weeks.OECs suspension[(1～1.4)?10 5 /ml ]was transplanted into transected spinal cord at T 10 level,while the DMEM solution was in ject-ed instead in the control group .At 6weeks and12weeks after transplantation,the rats were evaluated with cl imbing test and MEP monitoring.The samples of spinal cord were procured and st udied with histological and immunohistochemical measurement.Results At 6weeks after transplantation,all of the rats in both transplanted and control group were paraplegic,and MEPs could not be recorded.Morphol ogy of trans-plante d OECs was normal,and OECs were interfused with host well.Axons could regrow i nto gap tis sue be-tween the spinal cords.Both OECs and regrowed axons were im munoreactive for MBP.No re grown axons were found in the control group.At 12weeks after transplantation,2rats(2/7)had lower ex tremities mus-cle contraction,2rats(2/7)had hip and /or knee active movement,and MEP of 5rats(5/7)could be recorded in the calf in the transplantation group.Non e of the rates(7/7)in the control group had functional improvement,and n one of them had MEPs recorded.In the transplanted group,histological and imm unohis-tochemical methods showed the number of transplanted OECs reduced and s ome regrown axons had reached the end of transected spinal cord.However,no r egrown axons could be seen except scar forma tion in the control group.Concl usion Cryopreserved OECs could integrate with the host and promote re growing a x-ons across the transected spinal cord ends.

BACKGROUND:Olfactory ensheathing cells (OECs) have been shown to possess the potential of repairing injured spinal cord, but their biological characteristics after transplantation in vivo are not well understood.OBJECTIVE: To investigate the migration of OECs after transplantation into the injured spinal cord of adult rats.DESIGN: Randomized and controlled experiment.SETTING: Department of Orthopedics, Guangdong Provincial People's Hospital; Experimental Animal Center of the North Campus of Sun Yat-sen UniversityMATERIALS: Totally 38 2-month-old male SD rats with body mass of (350 ±20) g were used in this study.METHODS: This experiment was conducted in the Experimental Animal Center, North Campus of Sun Yat-sen University between February 2004and May 2004. Two SD rats were used to extract the OECs, which were stained with Hoechst 33342. Totally 36 SD rats were subsequently randomized into 3 groups, namely rostral transplantation group, caudal transplantation group and control group with 12 rats in each group. The rats in the rostral and caudal transplantation groups subjected to operations to establish thoracic spinal cord injury model and OEC suspension was injected; in the control group, the rats were spared of thoracic spinal cord injury with only OEC suspension injection.MAIN OUTCOME MEASURES: Distribution of OECs in the spinal cord was observed under fluorescence microscope 1, 2, 4 and 6 weeks after operation, respectively.RESULTS: Of the rats in the 3 groups, 1 died in the rostral group, and 2in each of the caudal transplantation and control groups, leaving 29 rats for result analysis. The OECs in the rostral and caudal transplantation groups migrated longitudinally along the long axis of the spinal cord to a farthest distance of 8 mm and penetrating the scar tissues, but very few cells could reach the contralateral side. The OECs in the control group diffused locally without migration.CONCLUSION: OECs mainly migrate along the axons in white matter of the injured spinal cord, and their rostral and caudal migration does not differ in speed or amount. Only a small amout of OECs can across the transected gap of the spinal cord.

Objective To study the relationship between the change in serum levels of vascular endothelial growth factor(VEGF) and endostatin (ES) after trascatheter arterial chemoembolization (TACE) and the prognosis of patients with liver carcinoma. Methods Serum VEGF and endostatin levels were measured by enzyme-linked immunoassay in 120 patients with hepatocellular carcinoma before and a week after TACE.Results Among patients with large (diameter ≥5 cm) tumors , the serum levels of ES and VEGF before and after TACE are 43.35 ( ±9.80),48.35 ( ± 10.89), 310.23 (±64.31) ,and 369.10 ( ±60. 11) ng/ml respectively. Among patients with portal vein tumor thrombus, the corresponding figures are 54.28 (±8.78 ),50.28 (±7.51), 331.26 (±63.38) and 400.29 (±60.98) ng/ml. The levels of VEGF and ES were significantly related to the presence of portal vein tumor thrombus, the clinicopathological grade and size of the tumor(P ＜0.05 ). Patients with a higher grade tumor were more likely to have elevated levels of VEGF and ES.So are patients with more advanced stage tumors. In addition, higher levels of VEGF and ES in serum are associated with worse survival. Conclusion Elevated serum VEGF and endostatin levels in patients with hepatocellular carcinoma are closely correlated with the grade and size of the tumor, and the presence of portal vein tumor thrombus. Serum VEGF and ES level may be used for predicting the biological behavior, invasion, metastasis and prognosis of hepatocellular carcinoma.

[Objective]This study was designed to construct a recombinant adenovirus vector contains LMP-1 gene,and investigate the osteoinductive activity of MSC which were transfected recombinated adenoviral vector carrying LMP-1 gene.[Methods]Total RNA was extracted from mt osteoblast and the LMP-1 gene was acquired by RT-PCR,the LMP-1 gene and entry vector pENTR/D-TOPO were used to create the entry clone with the directional TOPO clone technology,then the entry clone and the expression vector were used to create the expression clone throush the LR recombination reaction.The adenovirus expression clone was linearized by PacI and transfected to the 293A cell line to harvest a high titer.Ad-LMP-1 was infected into the 3rd passage MSC,the expression of LMP-1 was detected by Western blot.The osteogenic activity of MSC was evaluated by the expression of collagen Ⅰ,ALP,osteocalcin and the formation of bone nodule.[Result]The LMP-1 gene was successfully acquired and confirmed,the entry clone and the expression clone were both verified by enzymes digestion,and the expression clone was further confirmed by sequenced.The expression of LMP-1 was detected successfully in MSC.The increasing expression of collagen Ⅰ,osteocalcin.ALP and bone nodule were observed by comparing to the control group.[Conclusion]Gateway technology not only make construction of the pAd-LMP-1 recombination adenovirus vector simple and fast,but also get a high transfection efficacy in MSC.LMP-1 gene can induce the osteoblast differention of MSCs,and improve its osteogenic activity.The adenovirus vector is reliable to be used in further gene therapy research.

[Objective] To explore the techniques of hand-assisted laparoscopic splenectomy plus portozygos disconnection procedure. [Methods] 18 patients with portal hypertension were operated by hand-assisted laparoscopic splenectomy plus portozygos disconnection. [Results] The operations were completed successfully. The mean operative time was 180(150～260) minutes, the mean blood loss estimated was 540(200～2 000) mL, the mean weight of spleen was 910 (500～2000) g and the mean length of hospital stay was 9.7 (8～18) days after operation. All patients had recovered rapidly and suffered less pain and only 5 patients needed analgesic. The mean recovery time of the bowel function was 52(24～74) hours. The hand incisions healed well, and only one occurred complications and another suffered death. [ Conclusions ] Hand-assisted laparoscopic splenectomy plus portozygos disconnection procedure is to be feasible and safe, and has the merits of minimally invasive surgery.

Objectives:To investigate the influence of PN via portal vein on insulin and glucagon in liver regeneration. Methods:The rabbits were randomly devided into control group( n =5),PN via portal vein group(group Ⅰ, n =10) and PN via central vein group(group Ⅱ, n =10).The PN was performed for 6 days after partial hepatectomy.The concentration of serum insulin and glucagon in portal and perpheral vein were analysed with radioimmunoassay. Results:The concentration of serum insulin was increased in group Ⅰ and group Ⅱ,and it was increased significantly( P 0.05),but it was increased significantly in portal vein blood in group Ⅰ( P