Abstract: Molecular genetic map is a fundamental organizational tool for genomic research. However, a genetic linkage map for Bupleurum chinense DC. has not been developed. In this study, with the theory of pseudo-testcross, 96 F1 plants from an intraspecific cross of B. chinense were used as mapping populations. Twenty eight ISSR (inter-simple sequence repeat) primers and 44 SSR (simple sequence repeat) primers were used to detect the polymorphisms between the parental plants, and of them, 28 ISSRs and 14 SSRs were selected to analyze the F1 populations. The map consisted of 13 linkage groups which included 80 (72 ISSRs and 8 SSRs) loci, and covered 2 633.9 cM with an average density of 33.4 cM. All 13 linkage groups consisted of 2-31 loci ranging in length from 15.4-1295.7 cM. This map will provide a basis for studies on gene mapping, map-based cloning and maker-assisted selection of important traits in B. chinense.

The effects of vascular endothelial growth factor (VEGF) on neural differentiation of human embryonic stem cells (hESCs) in vitro and the possible mechanism were observed. The hESCs lines, TJMU1 and TJMU2, were established and stored by our laboratory. hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups: group A, routine induction; group B, routine induction+10 ng/mL VEGF; group C, routine induction+10 ng/mL VEGF+10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were detected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence. The percentage of Nestin positive cells in group B was significantly higher than in groups A and C, while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C (P<0.01). There was no significant difference between groups A and C (P>0.05). It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

Objective:To observe the nutritional status and respiratory function of the patients with mechanical ventilation in Intensive Care Unit after high fat and low carbohydrate enteral nutrition treatment.Methods:271 patients with mechanical ventilation in Intensive Care Unit were randomly divided into two groups:study group and control group.The study group was given high fat and low carbohydrate enteral nutrition treatment,while the control group with Enteral Nutritional Suspension.Thealbumin,prealbumin,blood gas analysis(PaC02,Pa0) and RQ,VC02,V02,IgG,CD4,CD4/CD8 were observed.Results:After nutritional support,nutritional status of two groups were improved,but there was no significant difference regarding the serum albumin and prealbumin.The PaO2,VO2,VCO2,RQ,IgG,CD4,CD4/CD8 and the total time of mechanical ventilation were improved,and the study group was superior to the control group.Conclusion:High fat and low carbohydrate enteral nutrition improves the nutritional status and respiratory function of the patients with mechanical ventilation in Intensive Care Unit,and decreases the total time of mechanical ventilation and total medical expenses.

This study was aimed to establish an identification method for Spatholobus suberectus and its adulterants by near-infrared spectroscopy (NIRS). Near-infrared diffuse reflection spectroscopy (NIRDRS) spectra of different S. suberectus and its adulterants were acquired by using OPUS INDENT analysis software. NIRDRS spectra clustering analysis model and identification model were established and verified. The results showed that S. suberectus from dif-ferent regions and its adulterants were identified successfully by clustering analysis model and identification model. It was concluded that Spatholobus suberectus and its adulterants can be identified rapidly and non-destructively by NIRS.

Objective To study the effects of urinary kallidinogenase (kallikrein) on focal cerebral blood flow (CBF) following cerebral infarction in rats by laser speckle imaging.Methods Permanent middle cerebral artery occlusion (MCAO) was induced in male Sprague-Dawley rats by the intraluminal filament technique.Laser speckle imaging was used to measure CBF in the ischemic cortical area and middle cerebral artery territory.The brain was stained with 2,3,5-triphenyltetrazolium chloride (TTC) to determine the infarct size.Neurological deficit score was measured.Results CBF increased in both hemispheric cortical area and MCA territory on the first and second days following urinary kallikrein administration at high dose but not at low dose.Larger blood vessel diameter and increased blood flow velocity were noticed in the high dose group in some arteries when compared to the low dose group and normal saline control group.At 36 h after cerebral ischemia,the brain infarct size was 10.14% ±3.02% ,25.99% ±3.90% and 27.10% ±3.32% in high, low dose and normal saline control groups,respectively.The infarct size was significantly smaller in the high ( F = 61.14, P<0.01 ) but not low dose group when compared to the normal saline control group.The neurological deficit was milder in the high dose group but not the other two groups at 4 h after cerebral ischemia; however, there were no differences among the groups at 36 h after MCAO.Conclusions Urinary kallidinogenase can reduce cerebral infarction volume and neurological deficit in rats following focal cerebral ischemia.These effects may be attributed to enhanced collateral circulation and improved CBF in the hemispheric cortical area and MCA territory.

Objective To evaluate a combination of open surgery and multiple interventional methods in the treatment of DVT of the lower extremities. Methods 521 cases (521 limbs,356 male and 165 female) were studied in this group. Age ranged from 16 to 86 years with the mean age of (46 ±9)years. All 521 cases with DVT were treated by Fogarty embolectomy catheter. Among them,348 cases underwent percutaneous transluminal angioplasty (PTA), 135 cases received PTA and ultrasound ablation,stent-grafts were implanted in 108 cases. Results Based on angiography during operation, the obstructed iliofemoral vein received complete recanalization in 511 cases. Among them, the postoperative luminal diameter was more than 90% in 38 cases after Fogarty embolectomy, the average stenosis rate was reduced from 90% ±5% to 24% ±5% in 365 cases after PTA and stent-grafts were implanted in 108 cases with the stenosis rate still over 50% after PTA. Only partial recanalization was achieved in the entrance of common iliac vein to inferior vena cava in 10 cases. Of the 521 cases,472 cases were followed-up with mean time of (53 ± 26) months, ranging from 8 to 108 months and 462 cases reported satisfactory results with normal life,the unsuccessful 10 cases still felt swelling pain especially in erect position. Complications occurred in 33 cases. Conclusions Open surgery combining with multiple interventional methods is a safe and effective method in the treatment of DVT.

Objective To evaluate the quality of Radix et Caulis Ilicis Asprellae from Pingyuan planting base and Chinese herbal medicine market. Methods The water- and alcohol-soluble extracts from 19 batches of Radix et Caulis Ilicis Asprellae medicinal materials were detected according to Appendix ⅨH, ⅩA of the Chinese Pharmacopoeia ( 2010 edition). And the quality of the medicinal materials was evaluated by microscopic identification technology according to the method for Radix et Caulis Ilicis Asprellae recorded in Guangdong Provincial Chinese Medicine Standard, and then thin layer chromatography ( TLC) was optimized to establish the high performance liquid chromatography (HPLC) fingerprint. The HPLC was performed on Waters XBridgeTM C18 column (250 mm × 4.6 mm, 5μm) with acetonitrile(A)-0.2% (v/v) phosphorus acid (B) as the mobile phase by gradient elution, flow rate was 1.0 mL/min, and detection wavelength was 220 nm. Results The results of sample characters, TLC and microscopic identification showed that the samples of Radix et Caulis Ilicis Asprellae in Chinese herbal medicine markets were certified products, but stems and roots were blended. Seven common peaks were showed by HPLC and confirmed by similarity analytical software. The similarity of 15 batches of planting base samples was all above 0.9. Of 19 batches of the commercial samples, the similarity of 11 batches was above 0.9. The alcohol-soluble extract contents were in the range of 64.55 mg/g to 186.18 mg/g. Conclusion The medicinal materials of Radix et Caulis Ilicis Asprellae from Chinese herbal medicine market are certified products, but the qualities vary greatly for the blending of stems and roots and inadequate growth years. The quality of materials from planting base is better. The established method is helpful for the quality evaluation and control of Radix et Caulis Ilicis Asprellae.

Objective To investigate the changes and significance of autophagy in rats with experimental acute necrosis pancreatitis (ANP).Methods According to method of random number,18 rats were randomly divided into control group,ANP group,ANP+rapamycin (RAP) group.The ANP rat model was established by intraperitoneal injection of 20％ L-arginine.The rats of ANP+RAP group were intraperitoneal injected with RAP 1.2 mg/kg at 30 minutes before modeling.The rats of control group were intraperitoneal injected with 0.9％ NaCl solution.The blood was drawed from the hearts nine hours after modeling for subsequent experiments.Serum levels of trypsinogen activation peptide (TAP),interleukin (IL-1),IL-6 and tumor necrosis factor (TNF) α were measured with enzyme-linked immunosorbent assay.The pancreatic tissues were pathologically scored.Autophagy-related structures in rat pancreatic acinar cells were observed by transmition electron microscopy.The expression of autophagy marker microtuble assciated protein 1 light chain 3 (LC3)-Ⅱ and Beclin-1 at mRNA and protein level were measured by quantitative real-time polymerase chain reaction (qRT-PCR),Western bloting and immunohistochemistry.The single factor analysis of variance was used for mean comparison among groups.Results A rat model of ANP was successfully established.Histopathological score of pancreas acinar cell necrosis of ANP+RAP group (2.19±1.38) was higher than that of ANP group (0.97±0.68),and the difference was statistically significant(F=33.75,P＜0.05).The results of Western blotting indicated that the protein expression of LC3-Ⅱ and Beclin-1 in ANP group (35.25±2.68 and 49.40±5.28)were higher than those in control group (1.54±0.16 and 0.78±0.06),furthermore the expressions in ANP+RAP group(123.53±3.21 and 76.41±3.80) were higher than those in ANP group,and the differences were statistically significant(F=2 045.54,326.87,both P＜0.01).Immunohistochemistry results also indicated that the LC3Ⅱ and Beclin-1 expression at protein level of ANP+RAP group (7 570.63±4 357.67 and 3 418.09±2 035.78) were higher than those of ANP group (1 926.53±1 414.44 and 536.11±403.10),and the differences were statistically significant (F=39.83,41.58,both P＜0.01).The expression of Beclin-1 at mRNA level of ANP group (107.12±29.10) was statistically higher than that of control group(7.01 ±3.39),and the difference was statistically significant (F=3.61,P＜0.05),but the expression of ANP+RAP group (97.63 ± 65.38)was no significant difference compared with ANP group.However,the expression of LC3-Ⅱ at mRNA level of ANP+ RAP group (4.37 ± 1.67) was statistically higher than that of ANP group (1.76 ± 1.59),and the difference was statistically significant(F=16.10,P＜0.05),but the expression of ANP group was no significant difference compared with control group (1.51 ±0.95).The result of electron microscopy showed that autophagy related structures increased in ANP group compared with that of control group,which of ANP+RAP group was more.The serum levels of TAP,IL-1 and IL-6 of ANP + RAP group were (36.47 ± 1.71) pmol/L,(122.88± 26.67) pg/mL and (107.39±13.95) pg/mL,which were all higher than those of ANP group ((25.63 ± 6.05) pmol/L,(98.06 ±9.29) pg/mL and (86.16± 7.20) pg/mL),and the differences were statistically significant (F=116.71,50.45,79.67; all P＜0.01).There was no significant difference in TNFα between ANP+ RAP group ((140.80±60.82) pg/mL) and ANP group ((105.23±6.95) pg/mL,F=14.76,P＞0.05).Conclusions Autophagy increased in rats with ANP.Promoting autophagy could significantly activate trypsinogen,aggravate pancreatic injury and increase inflammation reaction,which indicated that autophagy might involve in the pathogenesis of ANP through trypsinogen activation.

Objective To study the N terminal pro brain natriuretic peptide (NT-proBNP)in serum of hypertension patients. Methods 152 patients with hypertension were included in this study.According to the different degree and development of hypertension,152 patients were divided into three different grade group of hypertension:first grade of hypertension group (30 patients),second grade of hypertension group (36 patients)and third grade of hypertension group (86 patients).And the 86 patients were divided into hypertension group (40 patients)and hypertension with diabetes group (46 patients)re-spectively,comparaed with the level of NT-proBNP in serum of different hypertension grade groups.Results The levels of the NT-proBNP in serum were 68±44,122±31 and 834±309 pg/ml of first grade of hypertension group,second grade of hypertension group and third grade of hypertension group,respectively.The level of the NT-proBNP was gradually increased with grade of hypertension (t=2.455,3.561,P<0.01).The level of the NT-proBNP (1 178±864 pg/ml)of hypertension with diabetes group was significantly higher than hypertension group (599±411 pg/ml)(t=3.785,P<0.01).Conclusion The level of NT-proBNP can obj ectively reflect the grade of the disease in patients with hypertension.it is a certain signifi-cance guiding for monitoring the clinical treatment of the disease.

AIM: To investigate the protective effect and mechanisms of di-ao-xin-xue-kang (DK) on myocardial ischemia-reperfusion. METHODS: Models of myocardial ischemia-reperfusion were constructed with sixteen mongrel dogs. The left ventricular pressure (LVSP), the marximal/minimum rate of LVSP (?dp/dt max) and the serum aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA) were determined before and 90 minutes after occlusion, 120 and 240 minutes after reperfusion, and the MDA content in myocardial cell membrane prepared at 240 minutes after reperfusion was determined. The oxygen free radical was assayed with electron spin resonace spectroscopy (ESR) technique. RESULTS: ① LVSP and ?dp/dt max in the normal saline control (NS) group decreased with the time progress of occlusion-reperfusion, and it was the same in DK group, but the levels were significantly higher than that in NS group (P