Object To study the inhibitory effect and mechanism of extract from cultured cell of Taxus chinensis (Pilg.) Rehd. on hepatocarcinoma cell line SMMC 7721. Methods Trypan blue stain assay was used to determine tumor cell growth curve, flow cytometry and Giemsa staining were used to analyze cell cycle and cell apoptosis. Results Inhibitory effect happened on tumor cell line SMMC 7721, G 2/M stage of tumor cells increased with the addition of T. chinensis cell extract by carrene and apoptosis happened. Conclusion The extracts from cultured cell of T. chinensis have inhibitory effect on SMMC 7721 cells and can further induce cell apoptosis.

Objective:To investigate the sensitivities and specificities of high-frequency color doppler ultrasonography, thin layer CT enhancing scan and 99m Tc-MIBI biphasic scintigraphy in diagnosis of secondary hyperparathyroidism(SHPT)in uremia patients.Methods: High-frequency color doppler ultrasonography, thin layer CT enhancing scan and 99m Tc-MIBI biphasic scintigraphy were performed in 35 uric patients to detect the hyperplastic parathyroid glands. Then biopsies with thin needles were conducted with B ultrasound instruction in hyperplastic glands to determine the sensitivities and specificities of the above techniques and free combination of each 2 of them in diagnosis of SHPT. Results: The sensitivities /specificities of high-frequency color doppler ultrasonography, thin layer CT enhancing scan and 99m Tc-MIBI biphasic scintigraphy were 73%/95%, 75%/93% and 89%/98%, respectively; the sensitivity/specificity of 4 combinations: high-frequency color doppler ultrasonography plus CT, high-frequency color doppler ultrasonography plus 99m Tc-MIBI biphasic scintigraphy, CT plus 99m Tc-MIBI biphasic scintigraphy,and high-frequency color doppler ultrasonography plus CT plus 99m Tc-MIBI biphasic scintigraphy were 84%/90%, 93%/93%, 95%/93%, and 100%/90%, respectively. Conclusion: The sensitivity of 99m Tc-MIBI biphasic scintigraphy SHPT is higher than that of high-frequency color doppler ultrasonography and CT, but the specificity is not significantly different 3 photographic techniques. However, combination of high-frequency color doppler ultrasonography and 99m Tc-MIBI biphasic scintigraphy is simple and sensitive and can be regarded as the first choice for diagnosis of SHPT in uremia patients.

Objective:To optimize the overall desirability of orthogonal design sith multi index evaluations. Methods:Four multi index evaluations were applied to optimize the overall desirability of an orthogonal design,and their results wer analyzed. Results:According to the optimum conditions,there was no difference among the four multi index evaluations. Conclusion:multi index test can be used to represent the most desirable variables in experimental design.

Objective: To study the diagnostic characteristics of cardiac amyloidosis (CA) by non-invasive electrocardiography (ECG) in relevant patients. Methods: We retrospectively analyzed 60 CA patients diagnosed in our hospital from 2008-08 to 2013-12 for their clinical and ECG characteristics. Results: There were 48 male and 12 female patients with the ratio of 4: 1. The ifrst time diagnosis rate was low and the average age for conifrmed diagnosis was at (54. 5±14. 2) years.①There were 32 (53. 3%) cases combining heart failure, 12 (20%) with pleural effusion, 20 (33. 3%) with atrial arrhythmia, 8 (13. 3%)with ventricular arrhythmia, 4 (6. 7%)with sino-atrial block, 15 (25%)with atrio-ventricular block, 4 (6. 7%) with left bundle branch block (LBBB), 5 (8. 3%)with RBBB and 8 (13. 3%)with intra-ventricular block.②There were 32 (53. 3%) cases with low voltage on limb leads, 52 (86. 7%) with pseudo-infarct pattern, 48 (60%) with ST-T abnormality and 30 (50%) combining low voltage on limb leads with pseudo-infarct pattern.③The patients combining pleural effusion and with pseudo-infarct pattern had the increased ratio of low voltage on limb leads, while there were still 22 (45. 8%) cases without pleural effusion had low voltage on limb leads.④ ECG characteristics for 60 CA patients were as follows: QRS duration (104±26) ms, QT interval (404±34) ms, QTc (462±35) ms; the R wave of avR 0. 17 mV, QRS wave 0.30 mV; the R wave of limb leads and V1-3 were all<0.5mV, the S wave of V1-3 were 0. 62mV, 1. 61mV, 1. 56mV; the R/S ratio of V1-3 were 0. 19, 0. 12, 0. 20 respectively. Conclusion: CA patients had the highest incidence of pseudo-infarct pattern; meanwhile, combining with low voltage on limb leads, pseudo-infarct with long Q or S wave and ST-T abnormality but normal QRS duration was helpful for differential diagnosis of CA in clinical practice.

Objective To observe effects of Huanglian Jiedu Decoction on fasting blood glucose (FBG), fasting insulin (FINS), abdominal aortic morphology and expression of HIF-1α in the abdominal aorta in rats with type 2 diabetes. Methods The rat model of type 2 diabetes was established by intravenous injection of streptozotocin and feeding with high sugar and high fat diet. Rats were randomly divided into blank group, model group, control group and treatment group. The blank group and model group were given normal saline by gavage, the control group was given metformin, and the treatment group was given Huanglian Jiedu Decoction. Six weeks later, the levels of FBG and FINS were determined. Immunohistochemical method was used to determine the expression of HIF-1α in abdominal aorta and HE staining method to observe the changes of abdominal aortic morphology. Results Compared with model group and control group, abdominal aorta of HIF-1α and FBG level of treatment group were significantly decreased (P <0.01), FINS significantly increased (P <0.01), and abdominal aortic endothelial cell injury was significantly improved. Conclusion Huanglian Jiedu Decoction has a protective effect on the injury of vascular endothelium, and its mechanism may be related with inhibiting the expression of HIF-1α.

Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia (AA), high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ± 0. 29, 2. 26 ± 0. 37 and 1.58±0. 33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively (P < 0.01). The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0. 11,respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.

BACKGROUND: At present, the basic underlying molecular mechanism regulating the interactions among venous endothelial cells, platelets, leukocytes, and promoting local deep vein thrombosis microenvironment formation, still remains unclear, and there is no ideal method for early diagnosis of deep vein thrombosis. OBJECTIVE: To study the underlying role of nuclear factor kappa B1 and tissue factor in rats with deep vein thrombosis. METHODS: A total of 67 Sprague-Dawley rats were randomly divided into control group (n=10) and model group (n=57). Deep vein thrombosis model was established by a clamping and sewing method in femoral vein combined with cast fixation. The incidence and serious degree of thrombus were observed by dissecting rat femoral vein in different time points (2.5 and 25 hours after modeling). The model group was further divided into pre-thrombogenesis group (2.5 hours after modeling), thrombogenesis group (25 hours after modeling) and non-thrombogenesis group (25 hours after modeling). Then total RNA was extracted from the localized femoral venous endothelial tissue. The candidate genes, associated inflammation and thrombosis, were screened by a special gene chip. Then the gene expression of nuclear factor kappa B1 and tissue factor was further identified by real-time polymerase chain reaction. RESULTS AND CONCLUSION: Pre-thrombogenesis group had no thrombogenesis, while thrombogenesis group have 23 cases with thrombosis and non-thrombogenesis group have 22 cases without thrombosis. The results of gene chip hybridization analysis and real-time PCR found that the mRNA expression of nuclear factor kappa B1 and tissue factor in rat femoral vein endothelial tissue were significantly up-regulated at 2.5 hours after modeling (pre-thrombogenesis group was higher than control group) (P < 0.05), and continued up-regulating at 25 hours after modeling (thrombogenesis group was higher than the pre-thrombogenesis group, non-thrombogenesis group and control group) (P < 0.05). The results from present study indicate that up-regulating expressions of nuclear factor kappa B1 and tissue factor in local femoral venous endothelial tissue of rat deep vein thrombosis models may play a key role in initiating venous thrombosis.

BACKGROUND: The molecular mechanism and core regulatory network of deep vein thrombosis are not fully clarified yet.OBJECTIVE: To explore the roles of oxidative stress and Rac1/2 in rat deep vein thrombosis.METHODS: Deep vein thrombosis model in SD rats was established by a champing method femoral veins clamping combinedwith fixation of the lower extremity with plaster. The incidence and serious degree of thrombus were observed by dissecting ratfemoral vein at different time points (2.5 and 25 hours after modeling). The model rats were divided into pre-thrombogenesisgroup (2.5 hours after modeling), thrombogenesis group (25 hours after modeling) and non-thrombogenesis group (25 hours aftermodeling). Then total RNA and protein were extracted from the femoral venous wall tissues.RESULTS AND CONCLUSION: Colorimetry results showed that compared with the non-thrombogenesis group, theconcentration of malondiadehyde in rat femoral vein wall tissues of the thrombogenesis group was the highest (P < 0.05), followedby that of the pre-thrombogenesis group (P < 0.05). The concentrations of total superoxide dismutase and glutathione reductasein the thrombogenesis group were the lowest, followed by those in the pre-thrombogenesis group (P < 0.05). The results of genechip hybridization analysis and real-time PCR showed that compared with the non-thrombogenesis group, the expressions ofRac1 and Rac2 in rat femoral vein wall tissues of thrombogenesis group increased the most, followed by that of thepre-thrombogenesis group (P < 0.05). These findings indicate that the up-regulation of malondialdehyde and Rac1/2 as well asthe activity decrease of total superoxide dismutase and glutathione reductase may lead to the formation of deep venousthrombosis.

BACKGROUND: At present, the basic molecular etiological mechanism and core regulatory network of deep vein thrombosis (DVT) remains uncertain, and there is not an ideal measure for early diagnosis of DVT. OBJECTIVE: To study the underlying impact of cathepsin L/G in DVT rat model. METHODS: DVT rat models (n = 50) were established by clamping both femoral vein in three different positions within 3 seconds with mosquito forceps and fixing with cast. According to different observation phases and biological situations of the femoral vein thrombosis, model rats were divided into thrombogenesis group, pre-thrombogenesis group and non-thrombogenesis group. An additional 10 normal rats served as control group. Femoral vein was obtained at corresponding time points to exact total RNA. After a gene chip-based screening, the data of gene expression were further dissected by real-time PCR. RESULTS AND CONCLUSUON: Gene chip hybridization analysis results demonstrated that differential expression of cathepsin L/G gene was significant among groups, and the expression was greatest in the thrombogenesis group, followed by pre-thrombogenesis and non-thrombogenesis groups, which was significantly greater than the control group (P < 0.05). Real-time PCR analysis results were consistent with gene chip hybridization analysis results. These indicate that DVT is associated with an increase in expression of cathepsin L/G in local venous vascular wall, and they may be candidate molecular markers for early diagnosis of deep vein thrombosis.

Objective To explore the diagnosis value of morphology changes of pleomorphic megakaryocytes in the bone marrow (BM) smears and BM sections in chronic MPD(CML-CP, ET,PV and PMF). Methods BM aspiration was taken in 182 patients of MPD aspiration and biopsy examination was performed synchronously to obtain the BM smears and BM sections samples. The BM smears were subjected to Wright/Giemsa stain and immunohistochemistry stain, while the BM sections were subjected to Haematoxylin-Giemsa-Fuchsin stain. The morphology of pleomorphic megakaryocytes was classified into five groups, which were Ⅰ type ( inclusion type), Ⅱtype ( hypolobulated muclei type), Ⅲ type ( giant hyperlobulated nuclei type), IV type (micro pyknotic type), and V type(extrusion type). The size of megakaryocytes clusters was recorded as no clusters(0) , predominantly small clusters of fewer than 6 cells (1) or predominantly large clusters of at least 6 cells (2) . The detection rates of various types of pleomorphic megakaryocytes and megakaryocytes clusters were both analyzed in the BM smears and BM sections. Results In CML-CP group, the detection rates were (3. 73±3. 84)% , (14.19 ±7. 62)% ,(5.99 ±4.67)%, (34. 37 ±10.79)%, (9.45 ±6. 87)%, (32. 28 ±7. 67)% and 3.13 ±2. 30)% ,(12.61 ± 9.28)%,(4.94±4.27)%,(35.26±9.63)%,(9.47 ±5.89)%,(34.58 ±6.81)% for I tⅠype,Ⅱ type,Ⅲ type, Ⅳtype and Ⅴ type pleomorphic megakaryocyte in BM smears and BM sections. There were no significantly differences between the BM smears and BM sections(t value were 0.524,0.510,0.645, 0.239,0.011,0. 869,all P>0.05). In ET group, the detection rate of I type [ (6.17 ±2. 89)% ] in BM smears was significantly higher than that in BM sections [ 2.42 ± 1. 28) % ] (t = 7. 183, P < 0. 01) , while the detection rate of V type [ (6. 28 ± 3. 34) % ] in BM smears was significantly lower than that in BM sections [ (10. 18± 4.03) % ] (t = 3.940, P < 0.01). Besides these, the detection rates of other types were not significantly different between the BM smears and BM sections(t value were 0.079,0. 122,1.643, 1. 638,all P>0. 05). In PV group, the detection rate of V type in BM smears [ (6. 55 ±4. 11)% ] was significantly lower than that in BM sections [ (10. 30±3. 34) % ] (t = 2. 351, P < 0.05 ). However, the detection rates of the other types were not significantly different between the BM smears and BM sections (t value were 1. 635,0. 301,0. 132,0. 704,0. 681 ,all P' >0. 05). In PMF group, the detection rate of IV type in BM smears [(13.05 ±5.24)%] was significantly lower than that in BM sections [(29.14± 8. 72) % ] (t = 5. 245, P < 0. 01). And the detection rate of normal type in BM smears [ ( 33. 58 ± 14.39)% ] was significantly higher than that in BM sections [(23. 01±7.96)%] (t =2. 132,P<0. 05). Besides these, the detection rates of the other types were not significantly different between BM smears and BM sections( t value were 0. 787,0.646,2.062,0. 869, P > 0. 05 ) . In CML-CP and PV groups, the detection rates of size of clusters were not significantly different between the BM smears and BM sections (x~2 = 2. 772, P > 0. 05 ). In ET group, the detection rate of small clusters (1) in BM smears was obviously higher than that in BM sections, however, the detection rate of larger clusters (2) in BM smears was obviously lower than that in BM sections (x~2 = 13. 748, P < 0.01). In PMF group, the detection rate of no clusters(0) in BM smears was obviously higher than that in BM sections, however, the detection rate of large clusters(2) in BM smears was obviously lowers than that in BM sections (x~2 =18.741 ,P<0. 01). Conclusions Both BM smears and BM sections can be applied to observe pleomorphic megakaryocytes. The morphology changes of pleomorphic megakaryocytes have certain reference values for identification of MPD subtypes and differential diagnosis.