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The near infrared spectrum atlases of milk powders mingled with different adjuvant are the object for cluster analysis. Drawing assistance from the disparity in infrared fingerprint atlas that change according to the contents of chemical constituent, and making mingled component models, the milk powders mingled with different adjuvant were taken for a rapid sorting test using SIMCA clustering analytical method. In the experiment, two hundred fifty sorts of milk powders in the markets from different manufacturers were scanned by near infrared ray, and were tested with reproducibility determination. It was found difficult to extract fingerprint characters just from the external appearance of the near infrared spectrum atlases from milk powders mingled with different adjuvant, and it is needed to adopt pattern recognition technique to determine intelligently. One hundred sixty atlases were drawn out randomly for cluster analysis, and unknown samples were pretested. Results showed that the milk powders mingled with different adjuvant can be identified by near infrared spectrum analysis associated with cluster analysis methods, notwithstanding the similar near infrared spectrum atlases of different sample were difficult to identify directly. No overlapping phenomenon was found among milk powders mingled with different adjuvant, and they did not interfere with each other. The results from clustering spectra between samples were satisfactory, and the correct ratios of blind detections were over 90%. In addition, the correct ratios of this method may be elevated remarkably with sufficient number of samples, increasing training set sample quantity and sampling representation, and strengthening the standard degree of manipulation. It is concluded that the designed model to determine milk powders mingled with different adjuvant is rational, and the determination capability is fine.

This study was aimed to explore the apoptotic effect of Resveratrol (RES) on KG-1 cells and the expression of bcl-2/bax in vitro, and to clarify the possible mechanism of apoptotic effect of RES on leukemia cells. After treating with different concentrations of RES, the suppressive effect of RES on proliferation of KG-1 cells was analyzed by MTT method. Transmission electron microscope technique were used to detect the apoptosis status of KG-1 cells. The cell cycle and apoptosis percentage of KG-1 cells treated with RES were detected by flow cytometry. The expressions of bcl-2, bax mRNA and protein were assessed by semiquantitative RT-PCR and flow cytometry. The results showed that RES could obviously inhibit proliferation of KG-1 cells (p < 0.05). Compared with the control group, the cell number in S phase of KG-1 cells treated with RES increased (p < 0.05), the apoptosis rate enhanced significantly (p < 0.01) and the expression level was down-regulated, while expression level of bax was obviously up-regulated (p < 0.01). It is concluded that RES significantly induces apoptosis of KG-1 cells in vitro, which is probably related to the down-regulation of bcl-2 expression and up-regulation of bax expression.

Objective: To establish the qualitative and quantitative methods of Caratostigma willmottianum., Method: The roots of Radix Ceratostigmae were identified by TLC with the authentic medicinal material and plumbagin as the reference. The contents of plumbagin in Radix Ceratostigmae were determined by HPLC., Result: The TLC method was simple and specific. A Diamonsil C18 column was used. The mobile phase was methanol-water (75:25). Plumbagin in the sawple extract was separated well. The linear range of plumbagin was 33.6-313.6 ng. The average recovery of plumbagin was 100.3% and RSD was 1.9%., Conclusion: The methods can be used for identification and determination of plumbagin in C. willmottianum.

Objective: To study the technical conditions of the extraction and purification of the active composition from Polygonum orientale and Crataegus pinnatifida Bge. in Hongye Xingtong soft capsules with the macroporous resin., Method: The orientm, isorientm and hyperoside were used as index to screen the five kinds of resins. And the technical conditions of the enrichment and purification of D101 resin selected out of above were all-round studied., Result: The D101 was fit for adsorbing orientm, isorientm and hyperoside. Under the optimal conditions, the transfer rate of orientm, isorientm and hyperoside was above 91%, and the total solid was cut down by more than 60%., Conclusion: The D101 is greatly effective for the enrichment and purification of the active composition of P. orientale and C. pinnatifida Bge.

Sequential surface plasmon resonance imaging (SPRI) and imaging ellipsometry (IE) measurements are realized with a single set of optical components mounted on a goniometer. The sample situated on top of a triangular prism is positioned in the center of the goniometer. The resultant setup (SPRI/IE) can be used to examine the same surface region above (IE) and below (SPRI) the sample. Thickness values of silver stripes sputtered onto a gold substrate were determined by SPRI and IE, and the results were compared to establish the validity of the method. The SPRI/IE setup was also used to monitor the thickness of phospholipid films of different layer numbers. SPRI measurements were found to be more accurate for ultrathin films, whereas the IE results are more reliable for films whose thicknesses approach or exceed the distance encompassed by the evanescent wave of the surface plasmon. Thus, utilizing these two techniques sequentially facilitates the continuous monitoring of film thickness variation over a wide thickness range with high fidelity and provides a viable approach to image the surface regions with large topographic fluctuations.

Objective: To investigate vascular endothelial growth factor (VEGF) and its signaling pathway spontaneous response in type 2 diabetes mellitus (T2DM) mice to surgery-induced hind-limb ischemia., Methods: Sixty mice were randomly divided into two groups, one was fed with normal chow as control, and another was fed with high fat diet to induce T2DM. Fourteen weeks later, mice were surgically induced to hind-limb ischemia. Blood flow restoration was monitored with laser Doppler perfusion imaging. Tibialis anterior muscle was collected after 3 days of hind-limb ischemia. VEGF mRNA and protein expressions were analyzed using real-time PCR and ELISA; expressions of VEGF downstream signal molecules and receptors were analyzed using Western blotting and RT-PCR, respectively., Results: Perfusion recovery 10, 20, 30 days after ischemia was significantly attenuated in T2DM compared with control group (P < 0.05). T2DM impaired VEGF signaling pathway although VEGF levels increased in T2DM group. After ischemia, T2DM group had a comparable increase in VEGF expression compared with control group, but still had an impaired VEGF signaling pathway., Conclusion: VEGF signaling pathway is abnormal in T2DM mice, although VEGF had a response to the ischemic stimulation.

Objective: To investigate the perfusion restoration of type 1 diabetic mouse under the setting of surgically induced hind limb ischemia and the number and function of bone marrow endothelial progenitor cells (EPCs)., Methods: Forty mice were randomly divided into two groups: one group was injected with alloxan through tail vein to induce type 1 diabetes mellitus, and another group was set as control group. All mice were surgically induced to hind limb ischemia. Blood flow was monitored with Laser Doppler perfusion imaging for 4 weeks after artery ligation. Ten mice in each group were sacrificed and muscle tissues were harvested for histological detection. The remaining mice were sacrificed 7 days after surgery, bone marrow mesenchymal stem cells were harvested and EPCs were analyzed by flow cytometry and then were collected to culture for functional detection., Results: All mice received alloxan injection showed typical symptoms of type 1 diabetes mellitus. Restoration of blood flow was significantly slower in type 1 diabetic mice with lower level of vascular density in ischemic muscles than control group (P < 0.001, P < 0.05). The number and function of EPCs (CD34 and vascular endothelial growth factor receptor 2 double positive cells) in type 1 diabetic mice were significantly lower than that in control mice (P < 0.05)., Conclusions: The spontaneous angiogenesis is attenuated with a decreased number and function of EPCs in the setting of type 1 diabetes mellitus. This may partly explain why diabetic patients with peripheral artery diseases have more aggressive disease and poorer outcome.

Objective: To investigate the spontaneous response and its mechanism of type 2 diabetes mouse to surgical induced hind limb ischemia., Methods: Forty mice were randomly divided into two groups, one was fed with normal chow, and another was fed with high fat diet. Twelve weeks later, mice were surgical induced to hind limb ischemia. Blood flow was monitored. Four weeks later, mice were sacrificed and muscle tissues were harvested for histological detection, Western Blot, ELISA and RT-PCR examination respectively., Results: Spontaneous restoration of blood flow was significantly slow down in type 2 diabetic mice with lower level of vascular density in ischemic muscles. VEGF signaling pathway was impaired although the VEGF levels were increased in diabetes., Conclusions: The spontaneous response to ischemia was attenuated with an abnormal reaction of VEGF signaling pathway in the setting of type 2 diabetes mellitus.

Objective: To establish the fingerprint detecting standard of xinshao Injection by reversed phase high performance liquid chromatography., Method: The chromatographic conditions were as follows: an Diamonsil C18 column was used, the mobile phase was composed of 80% acetontrile and 0.1% phosphoric acid with gradient elution, the flow rate 1.0 mL min(-1), column temperature was 40 degrees and the UV absorbance detection was set at 230 nm., Result: A standard HPLC fingerprint procedure was developed for xinshao Injection, with 8 common peaks. Under the same selected chromatographic conditions, the HPLC fingerprints of raw drugs and their intermediate, finished products were obtained with good separation and correlation (no less than 0.97) according to the technical requirements of fingerprint on Injection of Chinese traditional medicine., Conclusion: This method was accurate, repeatable and can be used as the important parameters of the quality control for xinshao injection.

Objective: To investigate the prevalence of peripheral arterial disease (PAD) in China type 2 diabetic patients and to demonstrate the relationships between putative risk factors and PAD., Methods: In total 1,397 type 2 diabetic patients aged 50 years and older were enrolled and determined ankle-brachial index (ABI) and brachial-ankle pulse wave velocity (baPWV) in 15 Class III Grade A hospitals in 7 major cities of China., Results: Mean patient age was 63.7 +/- 8.2 years and mean duration of diabetes mellitus was 9.39 +/- 7.4 years. Two hundreds and seventy-two (19.47%) patients were diagnosed as PAD by ABI < 0.9, 122 (18.37%) in male and 150 (20.46%) in female. PAD patients had a significantly longer duration of diabetes mellitus, higher hemoglobin A1c, and a significantly lower mean body mass index than non-PAD ones. Aging, smoking, and systolic blood pressure were found to be positively related with the prevalence of PAD. In terms of lipid profiles, no variable was found to relate with PAD. Notably, baPWV showed as the same significant guiding index for PAD, almost matched with ABI., Conclusions: PAD is a common complication in China type 2 diabetic patients. Therefore, PAD screening and treatment should be emphasized for diabetic patients with high risk factors.

Objective: To search for a non-medicine therapy for infant emaciation., Methods: The observation group of 55 cases were treated with auricular point sticking combined with chiropractics. For the auricular point sticking therapy, vaccaria seeds were stuck at 9 points such as small intestine, Shenmen, spleen, endocrine, and others, and replaced once every 5 days, 10 days constituting one course. Chiropractics: knead or massage the muscles from lumbosacral area upward to Dazhui (GV 14). The control group were treated with oral administration of pepsin mixture, 10 mL after meals, thrice each day. They were treated for 5 courses., Results: Of the 55 cases in the observation group, 7 were cured, 20 markedly effective, 24 effective and 4 ineffective, with a total effective rate of 92.7%, and among the 23 cases in the control group, 2 cases were cured, 7 markedly effective, 8 effective and 6 ineffective, with a total effective rate of 73.9%., Conclusion: Auricular point sticking combined with chiropractics is a better non-medicine therapy for emaciation in children.

Objective: To assess the effectiveness and safety of non-permanent filter in preventing pulmonary embolism (PE) caused by deep venous thromboembolism (DVT)., Methods: The clinical data of 12 patients who were implanted with non-permanent filter for documented deep venous thromboembolism in Peking Union Medical College Hospital from September 2003 to June 2006 were retrospectively analyzed., Results: Among these 12 patients, 10 were implanted with temporary filters, and 2 with retrievable filters. All the 12 operations were successful without major complications. In the 10 patients who received temporary filters, filters were smoothly removed after 4 weeks in 9 patients, while one patient was further treated with anticoagulation therapy for 12 weeks because large emboli were entrapped by filter. In the 2 patients who received retrievable filters, filter were smoothly removed 2 weeks later in one patient; however, in another patient, the filter was left inside because large emboli were entrapped by filter. In all the 12 patients, emboli were entrapped in 6 filters (50%), in which the emboli were larger than 2 cm in 2 patients, larger than 1 cm but less than 2 cm in 1 patient, and less than 1 cm in 3 patients. PE scanning was performed in 10 patients before primary implantation, and PE was found in 5 patients. PE scanning was performed in 8 patients 6 months after implantation, and minor PE was found in 4 patients, whose symptoms and affected pulmonary artery were obviously improved. All patients received regular follow-up (ranged from 3 months to 2 years), 1 patient died of malignant tumor 4 months after operation, 1 patient suffered inferior vena cava occlusion due to large emboli entrapped by the temporary filter, and 1 patient experienced the recurrence of symptomatic DVT. Symptoms were improved in all the 5 patients with PE., Conclusions: Non-permanent filter can safely and effectively protect patients from PE. More standardized criteria for placement and protocols to ensure timely removal should be developed and implemented.

Objective: To evaluate the outcomes of percutaneous transluminal angioplasty (PTA) and stenting in the treatment of critical lower extremity ischemia of arteriosclerosis obliterans., Methods: From April 2005 to May 2006, PTA and stenting was performed on 29 limbs of 24 patients with critical lower limb ischemia. Among all these patients, their main clinical symptom was rest pain, while ulceration and gangrene were also found in 12 patients. Four limbs underwent PTA, 4 limbs underwent prosthetic femoral-popliteal bypass with infrapopliteal balloon dilatation simultaneously, and the remaining 21 limbs were performed by PTA and stent placement., Results: The success rate of endovascular procedure was 100% for femoral-popliteal artery, and 88% for infrapopliteal artery. Rest pain disappeared and the ulcers were healed after the operations. Only 1 patient received amputation during the operation., Conclusion: Endovascular therapy is an effective and safe procedure for the treatment of critical lower limb ischemia.

Objective: The specific signaling events by which circulating endothelial progenitor cells (EPCs) responding to alterations of the retinal vasculature induced by hypoxia are not yet completely understood. Our goal was to generate a suitable experimental model for the identification of such factors as well as the effects of these factors on the EPCs., Methods: Primary cultures of human retinal glial cells were incubated under hypoxic or normoxic conditions. Effects of hypoxia were confirmed by analysis of HIF-lalpha and VEGF expression. Circulating EPCs (CD34 +, CD133+ double-positive cells) were isolated by FACS Vantage from the human periphery blood. EPCs were treated with conditioned media from retinal glial cells incubated under either hypoxic or normoxic conditions. Effects of conditioned media on the proliferation and differentiation of EPCs were evaluated., Results: Incubation of human retinal glial cells at low oxygen concentration for 24 hours lead to HIF-lalpha expression and a significant increase of VEGF released to the media[ (319. 16 +/-34. 12) pg/ml versus (220. 28+/-24. 33) pg/ml] as compared to the control cultures incubated at normoxic conditions (P <0. 01). Treatment of EPCs with conditioned medium from retinal glial cells incubated at low oxygen concentration resulted in an increase of proliferative activity (day 3: 0. 83+/-0. 10 versus 0. 61+/-0. 22,P <0. 05; day 7: 1. 53+/-0. 21 versus 0. 88+/-0. 25, P <0. 05) and presence of differentiated endothelial cells as compared to cells cultured with conditioned media from normoxic cultures., Conclusions: Conditioned media from retinal glial cells contain factors capable of activating the proliferation and differentiation of circulating EPCs. Analysis of the effects of conditioned medium from neuronal glial cells incubated under hypoxic conditions on circulating human EPCs might be useful in providing an effective experimental method for the identification of the specific signaling molecules involved in the activation of circulating EPCs for the regeneration of retinal blood vessels.

Objective: To evaluate the effect of antisense monocyte chemotactic protein-1 (A-MCP-1) nanoparticles (NPs) as gene carrier on gene transfer in two kinds of animal models., Methods: Poly (lactic acid-co-glycolic acid) (PLGA) was used to make the NPs loaded with A-MCP-1 through a double-emulsion/solvent evaporation technique. NPs size was assessed by dynamic laser defractometer. The particle morphology was observed by scanning electron microscopy. DNA content in the NPs was measured by dissolving known amounts of NPs in chloroform and extracting DNA with water. In vitro release was performed in tris-EDTA buffer at 37 degrees C using double-chamber diffusion cells. The receiver buffer was replaced daily. The A-MCP-1 NPs was transfected into the cultured smooth muscle cells. PCR was used to evaluate the transfection of A-MCP-1. Cationic lipid (Lipofectamine) was used to transfect A-MCP-1 as control. After 48 hours incubation, cells were digested and examined by polymerase chain reaction. Twenty New Zealand white rabbits under jugular vein to artery bypass grafting procedure were divided into four groups: the first group received grafts treated with A-MCP-1 NPs, the second group received grafts treated with cationic liposome (dioleoyl trimethyl ammonium propane)-A-MCP-1, the third group received grafts treated with plasmid DNA, and the fourth group received grafts without transfection as control. Fourteen days after surgery the grafts were harvested. The expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blotting. The morphology of the grafts was investigated. To establish abdominal aortic aneurysms rats model, rats were randomly divided into three groups: A-MCP-1 NPs injection group, shame NPs injection group and control groups (without injection). Two weeks after surgery, diameter of abdominal aorta was measured and aortic tissue was obtained for PCR analysis to evaluate the A-MCP-1 expression. Western blot were applied to detect the inhibitory effect to the expression of MCP-1 mRNA and CD68 protein by A-MCP-1 NPs., Results: NPs size ranged 198nm to 205nm with average around 201.4 nm. DNA content in the NPs was 4.14%. NPs showed steady release rate in vitro in Tris-EDTA solution. It released faster in the first week then maintained a slowly sustained release up to 16 days. In cell culture A-MCP-1 gene successfully transfected into smooth muscle cells by NPs vector. In vein grafting animal model, A-MCP-1 expression was detected in the vascular walls of NPs and cationic lipid treated groups. The degree of vascular hyperplasia in the gene NPs treated group was significantly lower than that in control group. There was no significant difference in the inhibition of intimal hyperplasia between NPs and cationic lipid treated groups. Two weeks after transfection in abdominal aortic aneurysm rats models, the abdominal aortic diameter of A-MCP-1 NPs injection group was (1.79 +/- 0.12) mm, significantly smaller than that of control groups [shame NPs group was (2.58 +/- 0.21) mm, and saline group was (2.63 +/- 0.29) mm] (P < 0.01). The expressions of MCP-1 mRNA and CD68 protein in A-MCP-1 NPs injection group were 12.5 +/- 1.5 and 17.6 +/- 2.1, which were much lower than those in control group [in shame NPs group, which were 35.7 +/- 4.5, 42.3 +/- 5.7 (P < 0.01), and saline group which is 32.4 +/- 3.9, 39.8 +/- 4.8 (P < 0.01)]. Specific band of A-MCP-1 was detected only in the A-MCP-1 NPs injection group by PCR., Conclusion: A-MCP-1 gene NPs can be successfully used in rabbit vein grafting model and abdominal aortic aneurysm rats models, and may be potentially applied in clinical practice.

Objective: To determine the optimal strategies of occluded prosthetic bypass and to assess the risk factors which influence the outcome., Methods: From September 2003 to December 2004, 19 cases of failed low limb prosthetic graft were treated in the Peking Union Medical College hospital. All of them underwent operation, and the strategies of operation were determined by the cause assessed pre-procedure. Four cases underwent thrombectomy only, 5 cases underwent thrombectomy combined with the distal anastomosed-plasty, 3 cases underwent thrombectomy combined with profundaplasty, 2 cases underwent vascular re-construction with new prosthetic graft, 1 case underwent external iliac artery endarterectomy, 2 cases underwent stent cell transplantation, and the last 2 cases underwent amputated since the limbs showed un-salvageable. All the cases got at least 1 year following-up., Results: In the 17 vascular reconstruction cases except the 2 cases who underwent amputated, 14 cases (82.4%) got firstly successes, according to their limb ischemia symptom got well, but the rest 3 cases (17.6%) failed. And the mortality is 10.5% (2 cases). In the 1 year following-up, the prosthetic graft patency was 35.3% (6 cases), and the limb salvage was 76.4% (13 cases)., Conclusion: Optimal resolution for prosthetic graft re-occlusion should be based on good judgment of the occlusion cause. Then proper correcting the cause plus right following-up program could be the risk factors of prognosis.

Objective: To establish the qualitative and quantitative detective methods of Herba Polygoni Orientalis., Method: Isorientin, orientin, protocatechuic acid and gallic acid in Herba Polygoni Orientalis were identified by TLC. The contents of isorientin and orientin in Herba Polygoni Orientalis were determined by HPLC., Result: Isorientin, orientin, protocatechuic acid and gallic acid could be identified by TLC. Isorientin and orientin were well separated with Diamonsil C18 column and acetonitrile-0.1% phosphoric acid (18:82) as mobile phase. The linear range of isorientin was 0.075-0.90 microg. The average recovery of isorientin was 98.8% and RSD was 2.1%. The linear range of orientin was 0.041-0.49 microg. The average recovery of orientin was 98.8% and RSD was 2.1%., Conclusion: The methods can be used for qualitative identification and quantitation determination of Herba Polygoni Orientalis.

Objective: To evaluate the curative effect of the combination of endovenous laser treatment of varicose saphenous vein (ELVS) and classic ligation and stripping treatment., Methods: We retrospectively analyzed the clinical data of 21 patients with varicose saphenous vein (VS) who were treated with ELVS alone or combined with ligation and stripping in our hospital., Results: All the patients got good therapy result. The early symptom relief rate was 82.4%, while the late symptom relief rate was 100%. No infections, haematoma of wound, and any other major complications were reported. The common complications included pain induced by remains of the thrombosis phlebitis (n = 2, 11.7%), minor skin burn (n = 1, 5.9%), residue vein varicose (n = 1, 5.9%), numbness of the calf (n = 1, 5.9%), and mild peri-phlebitis (n = 1, 5.9%). All the complications were resolved after proper management. The 1-year follow-up showed no recurrence., Conclusion: The combination of ELVS and classic ligation and stripping is safe and effective in the treatment of varicose saphenous.

Objective: To study the chemical constituents of the essential substance from Polygonum oriental., Method: Compounds were isolated with silica gel and polyamide chromatography and their structures were determined by spectral analysis and chemical evidence., Result: Six compounds were obtained and identified as myricitrin, luteolin, gallic acid, catechin, protocatechuic acid, p-hydroxycinnamic acid., Conclusion: Six compounds were isolated from P. oriental for the first time.

Objective: To establish the fingerprint detecting standard of Compound Hongcao Injection by reversed phase high performance liquid chromatography., Method: The chromatographic conditions were as follows: an Inersil-ODS-3 column was used; the mobile phases (acetontrile and 0.1% phosphoric acid) with gradient elution; the flow rate 1.0 mL x min(-1) and the UV absorbance detection at 300 nm., Result: Under the same selected chromatographic conditions, the HPLC fingerprints of raw drugs and their intermediate, finished products were obtained with good separation and correlation according to the technical requirements of fingerprint on Injection of Chinese traditional medicine., Conclusion: 17 common peaks as well as their retention times and peak area ratios on HPLC fingerprint can be used as the important parameters of the quality control for Compound Hongcao Injection.

Objective: To establish a RP-HPLC method for the determination of gallic acid in Chinese herb Melastoma dodecandrum, and to evaluate the quality of the herb., Method: The RP-HPLC analysis was achieved by using a Polaris C18 column and tetrahydrofuran-methanol-0.2% phosphoric acid (0.5:0.5:99) as the mobile phase, with a flow rate of 1.0 mL x min(-1), and detected by UV at 274 nm., Result: The content of gallic acid was from 0.020% to 0.081%, in ten groups of M. dodecandrum collected from different locations., Conclusion: The method is a simple, convenient and rapid. It can be used for quality evaluation of M. dodecandrum.

Objective: To study the characteristics of cognitive dysfunction in rats with chronic cerebral ischemia and the effects of thyroid hormone on the rats' cognitive function., Methods: Thirty-one male SD rats were randomly allocated into normal control group (n=12), operation group (with bilateral carotid artery ligation and examined 5 weeks later, n=6), acute phase treatment group (APT, with bilateral carotid artery ligation and intragastric administration of thyroid hormone at 20 mg once daily for 5 weeks starting from the day of operation, n=7) and chronic phase treatment group (CPT, with the operation and thyroid hormone administration in an identical manner started from the sixth week following the operation, n=6). Morris water maze test was performed at the end of experiment. One-way ANOVA was used to estimate the differences in the learning and memory functions of the rats using SPSS10.0 for Windows., Results: The average latent period of the operation group was significantly prolonged in comparison with that of the other groups (P<0.05) in spatial orientation test. The probe time (time spent in the quadrant where the platform was once situated) of normal control was much shorter than those of the operation, APT and CPT groups in spatial probe test (P<0.05), and the operation group had the poorest score. The average latent period of the operation and CPT was longer than that of the other groups (P<0.05) in working-memory task (P<0.05), and the operation group again had the poorest score., Conclusion: Spatial cognitive function is totally damaged in rats with chronic cerebral ischemia, and learning can not induce the formation long-term memory because of short-term memory damage. Thyroid hormone may lessen but can not fully repair the damage of the cognitive dysfunction resulting from chronic cerebral ischemia, and early intervention with thyroid hormone may be beneficial for chronic cerebral ischemia.

Aim: To assess the contribution of signal transducer and activator of transcription 3 (JAK-STAT3) pathway, extracellular signal-regulated kinases1/2 (ERK1/2) pathway, and phosphatidylinositol 3-kinase (PI3-K) pathway to cardiomyocytes hypertrophy induced by cardiotrophin-1 (CT-1), a new member of interleukin-6 (IL-6) family of cytokines., Methods: STAT3, ERK1/2, and PI3-K were assessed by Western blot analysis. Activity of ERK1/2 was also confirmed by in-gel kinase assay. Hypertrophy of cardiomyocyte was evaluated by [3H]leucine incorporation and cellular protein-to-DNA ratio., Results: CT-1 simultaneously activated phosphorylation of STAT3, ERK1/2, and PI3-K in rat cardiomyocytes. Parthenolide, an inhibitor of STAT, suppressed CT-1-induced [3H]leucine incorporation by 88.3 % and protein-to-DNA ratio by 75.0 %. U0126, an MEK1/2 inhibitor, increased CT-1-induced the phosphorylation of STAT3 in a dose-dependent manner and, consistently, augmented CT-1-induced increase in [3H]leucine incorporation and cellular protein-to-DNA ratio by 17.6 % and 16.3 %, respectively. Wortmannin, a PI3-K inhibitor, did not influence CT-1-induced [3H]leucine incorporation and cellular protein-to-DNA ratio., Conclusion: The hypertrophic effect of CT-1 was essentially mediated by STAT3, independent of PI3-K, and negatively regulated by ERK1/2 via inhibiting the phosphorylation of STAT3. The interaction between STAT3 and ERK1/2 in CT-1-induced signaling contributes to development of cardiac hypertrophy.

Background: The Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway and the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway are the two major independent signal transduction pathways. However, it has recently been found that STAT3 may be negatively regulated by ERK1/2 in gp130-dependent signaling. Cardiotrophin-1 (CT-1), a potent novel hypertrophic cytokine, depends on gp130 to induce signaling and depends on STAT3 to exert hypertrophic effect. In this study, we examined whether STAT3 activity was negatively regulated by ERK1/2 during CT-1-induced signaling in rat cardiomyocytes and, if so, whether such crosstalk interfered with the hypertrophic effect of CT-1 and, furthermore, whether the mechanism underlying the crosstalk involved phosphorylation of serine 727 (S727) in STAT3., Methods: The activities of ERK1/2 and STAT3 were assessed by in-gel kinase assay and Western blot analysis, respectively. The role of S727 phosphorylation in the crosstalk between ERK1/2 and STAT3 was determined by a transient transfection study using a STAT3S727A mutant. Cardiomyocyte hypertrophy was evaluated by the cellular protein-to-DNA ratio and [(3)H]-leucine incorporation., Results: CT-1 simultaneously activated both ERK1/2 and STAT3 in rat cardiomyocytes. Inhibition of ERK1/2 by U0126 resulted in an increase of CT-1-induced tyrosine phosphorylation of STAT3 and, consequently, the protein-to-DNA ratio and [(3)H]-leucine incorporation. Transient transfection of the cells with STAT3S727A had no significant effect on CT-1-induced tyrosine phosphorylation of STAT3., Conclusions: STAT3 is activated by CT-1 in rat cardiomyocytes, but full activation is mitigated by the simultaneous activation of ERK1/2. The inhibition of ERK1/2 increases the activity of STAT3, which, in turn, enhances the hypertrophic effect of CT-1. The crosstalk between ERK1/2 and STAT3 is independent of the phosphorylation of the S727 in STAT3. Such crosstalk may contribute to the development of adequate cardiac hypertrophy.

Objective: To observe the dynamic changes of plasma inflammatory cytokine interleukin-1 beta (IL-1 beta) in patients with acute myocardiac infarction (AMI) before and after recanalization of the infarct related artery (IRA) and to observe the effect of recombinant human IL-1 receptor antagonist (rhIL-1ra) on the postischemic reperfused myocardium in experimental rabbits., Methods: (1) ELISA was used to measure the plasma IL-1 beta of 22 AMI patients, 20 males and 2 females, aged 64 +/- 12, before emergency percutaneous coronary intervention (PCI), and 12 hours and 24 hours after-intervention, and measure the plasma IL-1 beta of 8 healthy controls, 6 males and 2 females, aged 56 +/- 9. (2) Forty rabbits underwent ligation of the left circumflex branch of coronary artery (LCX) for 50 minutes and reperfusion for 4 hours after the ligatures were untied. The rabbits were randomly divided into 4 groups of 10 rabbits to be injected into the left ventricle immediately before the reperfusion with rhIL-1ra 10 mg/kg (group A), 20 mg/kg (group B), or 40 mg/kg (group C), and normal saline (control group) respectively. After reperfusion of 4 hours, the LCX was re-ligated. Evans blue was injected into the left ventricle. 15% KCl was injected intravenously to kill the rabbits. Their hearts were taken out to weigh the non-ischemic, ischemic, and necrotic cardiac muscles so as to calculate the infarct size. The myoperoxidase (MPO) activity was measured by colorimetry. Sections of myocardium were made. The number of apoptotic cardiomyocytes was evaluated by TUNEL method. The apoptotic rate of cardiomyocyte was measured by annexin V method. The DNA expression of myocardium was detected by DNA laddering method. The expressions of Bcl-2 and Bax apoptosis genes were assessed., Results: (1) The average plasma IL-1 beta level of the 22 AMI patients before emergency PCI was significantly higher than that of the controls (28 pg/ml +/- 9 pg/ml vs. 20 pg/ml +/- 11 pg/ml, P < 0.05), and became the highest 12 hours after the intervention (86 pg/ml +/- 14 pg/ml), and the high level lasted to 24 hours after emergency PCI. (2) In the ischemia-reperfusion rabbit model, the infarct size was 47% +/- 7% in the group A, 34% +/- 8% in the group B, 31% +/- 6% in the group C, and 61% +/- 11% in the control group (P < 0.05, 0.01, and 0.01 respectively). The activity of myocardial MPO was 16.6 +/- 3.6 min(-1).g.w.w(-1) in the group A, 10.9 min(-1).g.w.w(-1) +/- 1.9 min(-1).g.w.w(-1) in the group B, 7.8 min(-1).g.w.w(-1) +/- 2.2 min(-1).g.w.w(-1) in the group C, and 20.5 min(-1).g.w.w(-1) +/- 4.5 min(-1).g.w.w(-1) in the control group (P < 0.05, 0.01, and 0.01 respectively). The cardiomyocyte apoptosis evaluated by TUNEL was 38.3 n/HP +/- 7.4 n/HP in the group A, 25.6 n/HP +/- 6.8 n/HP in the group B, 12.2 n/HP +/- 3.3 n/HP in the group C, and 44.4 n/HP +/- 9.5 n/HP in the control group (P < 0.05, and P < 0.01 respectively in comparison between the group B and the control group and between the group C and the control group). The apoptotic rate by annexin V method was 11.6% +/- 2.7% in the group A; 7.7% +/- 2.4% in the group B, 4.7% +/- 1.4% in the group C, and 15.6% +/- 3.5% in the control group (P < 0.05, 0.01, and 0.01 respectively). DNA electrophoresis showed scaling ladder pattern only in the control group. The fluorescent density of the apoptosis gene Bax in myocardium was 24.9 +/- 8.2 in the group A; 15.5 +/- 3.4 in the group B, 10.6 +/- 2.5 in the group C, and 33.3 +/- 9.4 in the control group (P = 0.0298, 0091, and 0052 respectively) and no significant difference in the expression of Bcl-2 was shown among the 4 groups. Myocardial MPO was correlated with cardiomyocyte apoptosis (r = 0.86 by TUNEL, P < 0.01; r = 0.75 by Annexin V method, P < 0.05)., Conclusion: Inflammatory cytokine IL-1 beta is involved in myocardial ischemia-reperfusion injury. With potential therapeutic value in prevention and treatment of ischemia-reperfusion injury to myocardium, rhIL-1ra may reduce myocardial ischemia-reperfusion injury by suppression of cardiomyocytes apoptosis mediated by IL-als; 0.86 by TUNEL, P < 0.01; r = 0.75 by Annexin V method, P < 0.05)., Conclusion: Inflammatory cytokine IL-1 beta is involved in myocardial ischemia-reperfusion injury. With potential therapeutic value in prevention and treatment of ischemia-reperfusion injury to myocardium, rhIL-1ra may reduce myocardial ischemia-reperfusion injury by suppression of cardiomyocytes apoptosis mediated by IL-1.

Objective: To observe the dynamic fluctuation of inflammatory cytokines (IL-1beta, TNFalpha) and anti-inflammatory cytokine (IL-10) in acute myocardial infarct (AMI) patients before and after recanalization of infarct related artery (IRA) and analyze the relationship between fluctuation of cytokines and reperfusion state of myocardial tissue., Methods: (1) In 22 AMI patients and 8 healthy subjects, plasma IL-1beta, TNFalpha and IL-10 were measured with ELISA before emergency percutaneous coronary interposition (PCI), 12 h and 24 h post-intervention. (2) The 22 AMI patients were further divided into 2 groups according to ST-segment change in ECG at 2h after reperfusion: group A, good reperfusion of myocardium, ST decrease >or= 70% (n = 12) and group B, poor reperfusion of myocardium, ST decrease < 70% (n = 10). The change of plasma levels of IL-1beta, TNFalpha and IL-10 of the two groups was compared., Results: (1) Plasma TNFalpha and IL-10 in group A and B were not significantly higher those that the healthy controls (P > 0.05) before emergency PCI, but IL-1beta was significantly higher (P < 0.05). Plasma IL-1beta and TNFalpha in group A and B and IL-10 in group B at 12 h and 24 h post-intervention were significantly higher than those before PCI (P < 0.01, P < 0.05, P < 0.05), while IL-10 in group A was not (P > 0.05). (2) Plasma IL-1beta, TNFalpha and IL-10 in group B were not higher than those in group A before PCI, but they were significantly higher at 12 h post-PCI (P < 0.01, P < 0.05, P < 0.05) and IL-1beta and IL-10 were still higher at 24 h post-PCI (P < 0.05). The increment of IL-10 was significantly less than that of IL-1beta and TNFalpha in group A and B (P < 0.01, P < 0.05)., Conclusions: Cytokines may be involved in myocardial ischemia-reperfusion injury and increase of inflammatory cytokines may be a marker of poor myocardial microcirculatory reperfusion.

Objective: To experimentally investigate direct intramuscular gene transfer of nanoparticles encoding vascular endothelial growth factor for the treatment of peripheral artery disease., Methods: The human VEGF(165) cDNA was cloned into the eukaryotic expression vector PIRES2 under the control of cytomegalovirus promoter/enhancer. The recombinant gene was transferred into a rabbit model of chronic hindlimb ischemia by naked plasmid and nanoparticle respectively. Ischemia was induced in the hindlimb of New Zealand White rabbits by ligation of the distal external iliac artery and complete excision of the femoral artery and all its branches. At day 7 postoperation animals received VEGF(165) plasmid (10 intramuscular) or nanoparticle-VEGF(165) (8 intramuscular). With RT-PCR, immunohistochemistry analysis, and angiography, the expression and biological effects of VEGF(165) gene in experimental animals were investigated., Results: Two weeks after initiation of therapy, angiography showed that the transfer of VEGF(165) gene stimulated the formation of focal neovessels and established collateral circulation. The adductor muscle of ischemic limbs was histologically examined at day 14. Capillary density was increased among VEGF(165)-transfected rabbits, especially Nano-VEGF(165)-treated animals (Naked VEGF(165) plasmid = 50.18 per mm(2), Nano-VEGF(165) = 81.22 per mm(2), Control = 29.54 per mm(2), P < 0.05). RT-PCR showed that the transcription and expression of VEGF(165) gene in experimental group were significantly higher than those of control groups., Conclusions: Intramuscular administration of VEGF(165) induces collateral artery augmentation in the rabbit model of chronic limb ischemia. Nanoparticle can act as a vector to transfect specific gene and it will benefit gene transfer.

Objective: To discuss the diagnosis and surgical treatment of carotid body tumour., Methods: Thirty-eight cases (41 carotid body tumours) admitted from 1983 to 2002 were analyzed retrospectively., Results: All patients were examined by ultrasound and angiography routinely before operation. The diagnostic coincidence rates of ultrasound and angiography are 95% and 98% respectively. Thirty-eight tumours were resected successfully. Twelve patients underwent external carotid artery severed, internal carotid artery repaired and tumour resected operation. Eleven patients underwent external carotid artery severed and tumour resection. Eight patients underwent tumour simple resected operation. Six patients underwent external carotid artery severed and internal carotid artery reconstruction. One patient underwent tumour resection, while common, external, and internal carotid artery were partially resected., Conclusions: For suspected carotid body tumour patients, ultrasound and angiography reconstruction of carotid artery should be applied routinely before operation. The early stage and one stage tumour resected operation as well as internal carotid artery reconstruction are the key factors of decreasing operative complication rate.

Objective: To establish a method of quality control on qianliebeixi capsules., Method: To clarify semen vaccarine by TLC, to determinate the contents of quertin and kaempferol in prepations, by HPLC. Chromatographic conditions: C18 column with METH-0.2%H3PO4(47:53) as mobile phase; the temperature of column was 40 degrees C, and quercetin was detected at 370 nm with flowing rate 1 ml/min., Result: The spots of semen vaccarine were clear in TLC, no interference in negative contrast. The average recovery rate of quertin was 98.9% and the RSD was 2.2%; the average recovery rate in kaempferol was 100.4% and the RSD was 1.7%., Conclusion: The resolution of spots were perfect in TLC, the spots were easy to distinguish, The method is simple, rapid as well as reproducible and can control the quality of qianliebeixi capsules effectively.

Objective: To evaluate the diagnosis and treatment of mesenteric venous thrombosis., Methods: The clinical data of 11 cases diagnosed as mesenteric venous thrombosis between 1992 and 2001 in PUMC Hospital were analyzed retrospectively., Results: Postoperative state(27.3%), especially cirrhosis and portal hypertension, and other history of thrombosis (27.3%) were the most common causes. Thrombolysis was performed successfully in two of the eleven cases. The rest of them were misdiagnosed in other hospitals and operated. No patient died after operation, and one (11.1%) recurrence was found., Conclusions: Early application of anticoagulant is necessary for patients with thrombosis risks. For suspected patients, early computed tomography (CT) and DSA examination should be performed, and prompt thrombolysis and anticoagulation therapy can be performed to avoid the bowel resection after definite diagnosis. To reduce the recurrence, anticoagulant should be maintained for a proper time.

The genetic diversity of fifty-six indigenous pig breeds in China, and three introduced pig breeds (Duroc, Landrace and Large White) was surveyed using twenty-seven microsatellites recommended by the International Society for Animal Genetics (IS-AG) and Food and Agriculture Organization (FAO). By means of the allele frequencies, mean heterozygosity, effective number of alleles, estimator of gene differentiation, polymorphism information content, genetic distance and dendrogram analysis, the genetic variability and population structure of native pig breeds were estimated. Genetic variabilities within native pig breeds are as follows: Effective number of alleles vary from 2.12 to 9.03, from 0.44 to 0.87 for mean heterozygosity, from 0.39 to 0.86 for polymorphism information content. Nei's genetic distance and Nei's standard genetic distance were estimated and used to construct UPGMA and NJ dendrograms, which were evaluated by the bootstrap test. Fifty-six Chinese indigenous pig breeds were clustered into twelve groups based on the dendrogram. Compared with the classification in Pig Breeds in China, I, II and III groups in the study are equivalent to North-China type, IV group basically accords with Lower-Changjiang River Basin type, V, VI, VII, VIII and IX groups quite correspond with Central-China type, X and XI group largely correspond to South-China type, the last group, XII is equal to South-west type at large. Suggestions that the conservation farms together with conservation areas are appropriate methods for the preservation of native pig breeds in our country were proposed. The results could provide basic molecular data for the research on the germplasm characteristics of local breeds in our country and scientific basis for the conservation and utilization of those breeds.

An efficient method with microchemical method and solid phase extraction (SPE) technique for the determination of oxadiazon residues in cereals by gas chromatography-mass spectrometry (GC-MS) has been established. The oxadiazon was extracted from the sample with benzene-hexane(1:1, V/V). The extract was cleaned-up by a column of 500 mg Supelclean LC-Alumina-N. When the blank rice and corn were spiked with oxadiazon in the range of 0.005 mg/kg-5.00 mg/kg, the recoveries were 90.4%-115.7% for rice and 81.3-109.7% for corn, and with RSD of 2%-6% and 4%-9% respectively. The limit of detection was 0.005 mg/kg. The method is rapid, sensitive, accurate and suitable for the analysis of pesticide residues.

The virulence and drug susceptibility of a clinical isolate of Mycobacterium celatum which showed smooth transparent (ST) and smooth opaque (SO) colonies were studied. While ST cells multiplied intracellularly and maintained their coccobacillary form in a human macrophage model of infection, SO cells formed long filaments and completely destroyed the phagocytes. In BALB/c mice, the ST variant, but not the SO variant, grew efficiently in the spleen, liver and lung. The ST variant was usually more resistant in vitro than the SO variant to drugs, with MIC values for clarithromycin (CLA), azithromycin (AZI), ciprofloxacin, sparfloxacin, amikacin, clofazimine, ethambutol and isoniazid being higher than those of the SO variant. In beige mice infected with the more highly virulent variant ST, CLA and AZI were the most active drugs in terms of viable count reduction in organs and mutant selection. Together, these observations indicate that the ST variant of M. celatum is a virulent form that can be efficiently inhibited in vivo by CLA and AZI.