AIM: To establish a HPLC method for simultaneously determining chlorogenic acid,caffeic acid,forsythin,forsythoside A and baicalin in Shuanghuanglian Powder for Injection (Flos Lonicerae japonicae,Fructus Forsythiae,Radix Scutellariae,etc.).METHODS: HPLC assay was carried out on a Phenomenex~((R)) Luna C_(18) column (250 mm×4.6 mm,5 μm).The mobile phase was methanol-0.2% phosphoric acid solution as gradient elution.The flow rate was 1.0 mL/min.The detection wavelength was set as multi-wavelength detection.RESULTS: The linear ranges of chlorogenic acid,caffeic acid,forsythin,forsythoside A and baicalin fell within the range of 0.204-2.04 μg ,0.010 2-0.102 μg,0.186-1.86 μg,0.058 5-0.585 μg and 2.52-25.2 μg,respectively.The five components showed good linear correlations ( r = 0.999 7,0.999 4,0.999 8,0.999 8,0.999 9 ).The average recoveries of five components were in accordance with the determination requirements.CONCLUSION: The method is simple,quick,reliable,accurate and very suitable to the quality control of Shuanghuanglian Powder for Injection.

Objective To observe the effects of acupuncture combined with traditional Chinese medicine on scores of TCM syndrome and Neck Pain Questionnaire (NPQ) in patients of cervical spondylosis with syndrome of cold locking the collaterals. Methods Ninety cases of patients were randomly divided into acupuncture combined with TCM group, TCM group, and acupuncture group, 30 cases in each group. The acupuncture combined with TCM group was given modified Guizhi plus Gegen decoction and electro acupuncture;the TCM group was given modified Guizhi plus Gegen decoction only;the acupuncture group was given electro acupuncture only. The scores of TCM syndrome and NPQ of the three groups were observed and analyzed comparatively. Results The deviation scores of TCM syndrome and NPQ before and after treatment of the acupuncture combined with TCM group were higher than the acupuncture group and the TCM group (P<0.05), but there was no statistical difference between the acupuncture group and the TCM group (P>0.05). Conclusion Acupuncture combined with traditional Chinese medicine can relieve clinical symptoms of patients with cervical spondylosis with syndrome of cold locking the collaterals, ease their cervical pain, and improve their cervical function.

Objective To determine the total sodium in Danshen for injection.Methods Flame atomic absorption spectroscopy was used to determine the total sodium in Danshen for injection.Results The calibration curves were linear in the range of 0.5～10 mg/L for sodium chloride.The average recovery rates of sodium chloride were 101.8%,RSD=1.60%.Conclusion This method was found to be simple,quick and accurate for simultaneous determination of sodium in Danshen for injection.

The evaluation system of PBL teaching was constructed to develop PBL teaching in combination with the characteristics of medical courses. It covered students evaluation system, teacher's evaluation, and teaching pattern system in all aspects of teaching. The present study was carried out through repeated verification in many clinical medicine curriculum and many grades of students, and standardized assessment model, a more mature evaluation system, has been fully applied. All the indices of the evaluation system were comprehensive in contents, scientific, objective, manipulated easily and with good results, which is worth promoting the application of PBL teaching in the future.

Aim To explore the effect of exogenous nerve growth factor(NGF) on tau hyperphosphorylation in rat hippocampus during focal cerebral ischemia/reperfusion(I/R).Methods Focal cerebral ischemi-a/reperfusion model was induced by occlusion of the right middle cerebral artery using the intraluminal suture method.The level of tau hyperphosphorylation at Ser199/202 site and total tau in rat ispolateral hippocampal CA1 regions during focal cerebral ischemia/reperfusion were detected by SABC immunohistochemical method and Western blot,and the positive products were analyzed by image analysis system.Results The level of tau hyperphosphorylation at Ser199/202 site and total tau in rat hippocampal CA1 regions were significantly higher in I/R group than that in sham group(P

BACKGROUND: Due to the source of conspecific heart transplanted organ is limited and it becomes more and more deficient, and porcine cardiovascular system and haemodynamics are very similar to that of human being, therefore, domestic and foreign scholars have paid more attention on the heterogeneic heart transplantation from swine to human, and there are more and more basic studies about it.OBJECTIVE: To compare the zero-stress state of ascending aorta and pulmonary artery trunk between swine of different months and healthy persons, which provides necessary vascular tissue reconstruction basis for anastomosing vessel of heterogeneic heart transplantation from swine to human being.DESIGN: Open experiment.SETTING: Laboratory of Biomechanics, Yunyang Medical College.MATERIALS: This experiment was carried out at the Laboratory of Biomechanics, Yunyang Medical College from October 2001 to September 2002. Human heart specimens were chosen from 6 male adult corpses who had no visible manifestation of cardiovascular diseases, aged 18 to 30 years. Informed consents were obtained from all the relatives. Totally 42 conspecific swine, aged one month, were also chosen and divided into 7 groups, named as 1,2,3,4,5,6 and ＞ 6 months groups respectively, with 6 in each group. Swine were anesthetized at 1,2,3,4,5,6,＞ 6 months respectively and sacrificed in each group.METHODS: The corresponding ascending aorta and pulmonary artery trunk were dissected and isolated. Each arterial anterior wall was labeled in situ with coloring pen, and then artery was shifted into the oxygen-saturated Kreb's solution. Five isometric vascular rings were divided along the axial direction, then each vascular ring was cut open along the radial direction. Opening angle of each vascular ring at zerostress state was measured and recorded by computer 20 minutes later(The included angel formed through connecting the middle point of inner wall of open vascular plane and two end points of inner wall). The experiment was conducted at room temperature 20-30 ℃. All the procedures of vascular samples were performed within 24 hours after subjects died.MAIN OUTCOME MEASURES: Measuring results of opening angle of ascending aorta and pulmonary trunk at zero-stress state of swine at different months and human being.RESULTS: ①Comparison of measuring results of opening angle of ascending aorta and pulmonary trunk between swine at different months and human being: Opening angle of ascending aorta of human being increased from (100.4±12.8)°at proximal end to (152.8±18.6)°at distal end. Opening angle of swine at different months increased from about 68°at proximal end to about 130°at distal end; Opening angel of pulmonary artery trunk of human being did not changed significantly along axial direction, and opening angel of pulmonary artery trunk of swine at different months increased from about 65° at proximal end to about 125°at distal end. There was significant difference of opening angle of ascending aorta and pulmonary trunk at zero-stress state between human being and swine (P=0.028 ＜ 0.05).But the opening angle of ascending aorta of human being was similar at the second segment to at the fourth and fifth segments of swine of different ages; Opening angle of each segment of pulmonary artery trunk of human being was close to that of proximal end (first and second segments) ofpulmonary artery trunk of swine.② Comparison of measuring results of ascending aorta and pulmonary artery trunk at zero-stress state of swine at different months: Opening angle of ascending aorta and pulmonary artery trunk of swine did not changed with the increase of age (P=-0.063 ＞ 0.05).CONCLUSION: Opening angels of ascending aorta and pulmonary artery trunk at zero-stress state are close in some segments between human being and swine, suggesting that corresponding ascending aorta or pulmonary artery trunk can mutually anastomose in some segments in the heterogeneic heart transplantation from swine to human, and the effect of age is not significant in vascular reconstruction.

BACKGROUND: So far it has not achieved satisfactory effects to use synthetic blood vessels (diameter＜ 6 mm) as the substitutes for human small arteries or veins when clinical reconstructive vascular operations are performed. So, study on the substitutes for small arteries and veins has been a problem demanding prompt solution at present.OBJECTIVE: To observe the pressure-volume (P-V) relationship of umbilical cord vein of different gestational ages so as to provide theoretical basis of biomechanical properties for the clinical application of human umbilical cord vein as material for vascular grafting.DESIGN: Non-randomized controlled experiment.PARTICIPANTS: The experiment was completed in the Laboratory of Medical Biomechanics, Yunyang Medical College in June 2004. Umbilical cord veins of 50 normal fetuses were collected on spontaneous miscarriage or labor with the pregnant women's permission by the Department of Obstetrics and Gynecology, Shiyan Taihe Hospital. The gestational age of the fetuses ranged from 24 to 42 weeks and the pregnant women aged 20to 30 years. Among them, 8 cases with gestational age of 24-27 weeks,7cases 28-32 weeks,8 cases 33-36 weeks,4 cases 37 weeks,5 cases 38weeks, 5 cases 39 weeks , 5 cases 40 weeks , 4 cases 41 weeks and 4cases 42 weeks.INTERVENTIONS:Umbilical cord veins of 50 normal fetuses were collected. A 2-cm long segment of each umbilical cord was measured and labeled at both ends. Then they were cut off and put into normal saline (NS). The umbilical veins were carefully stripped from the cord; its two ends were fixed on the biomechanical experiment stand of soft tissues. The P-V relationship of fetal umbilical cord vein was measured, and then the compliance calculated.MAIN OUTCOME MEASURES: Pressure, volume and compliance of different umbilical cord veincurve of pressure-volume relationship of the umbilical cord vein was similar at week 37 to 40 but decreased significantly at week 28 and week 42. After regression analysis twice, it was found that the absolute value of regression vein increased with the increasing of gestational age[24-27 weeks (2.22±0.34)cord vein collected at 37 to 40 weeks of gestational age was similar. When the umbilical cord veins older than 42 weeks or under 28 weeks were compared, there was significant difference in their compliance (F=65.84-86.52, P＜ 0.01).CONCLUSION: Human umbilical cord veins are good substitutes for The relationship between compliance and gestational age. Our studies suggest that the umbilical cord vein, whose gestational ages range from 37to 40 weeks, is a material of choice for the transplantation.

In order to provide morphological data and theoretical basis for pig-to-human hepatic xenotransplantation, the difference in morphological parameters and vessel wall structural factors between human and porcine hepatic portal vein was studied. From human subjects and pigs of varying ages, hepatic portal veins were collected, paraffin-embedded and cut into sections. The histological structures were stained with HE, and elastin, collagen and smooth muscles were stained with Weigert, Aniline blue and orange G, respectively. Morphological parameters and relative contents of structural components were determined under microscopy and by computer image analysis system, respectively. The results showed that histological structures of human and porcine hepatic portal vein wall were similar. Caliber, wall thickness, lumen and wall area in pigs increased with age, all in linear correlation to months. Morphological parameters of 6- month-old pigs were similar to those of human. In pigs, collagen content increased gradually with months, elastin content remained relatively stable, smooth muscle content reached the peak at the 3rd month, and collagen/elastin (C/E) rose gradually. The contents of collagen and elastin in porcine hepatic portal vein wall were lower, while the content of smooth muscle was higher than in human, and C/E at the 5th and 6th month was similar to that in human. It is concluded that morphological parameters and contents of structural components of porcine hepatic portal vein vary with age. At the 6 month, its caliber, wall thickness, lumen and wall area are similar to those of human. There are differences in contents of structural components between human and pigs. However, in terms of C/E, mechanic properties of pigs at the 5th and 6th month mimic those of human, hence inosculation is viable in xenotrans-plantation between pigs and human.
Collagen/*analysis ; Elastin/*analysis ; Image Processing, Computer-Assisted ; Liver Transplantation ; Muscle, Smooth, Vascular/cytology ; Portal Vein/*anatomy & histology ; Portal Vein/chemistry ; Swine ; Transplantation, Heterologous

Objective To study the influence of pulsed electromagnetic fields on the expression of type I collagen by bone marrow mesenchymal stem cells and it's mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro.The third passage cells were harvested and exposed to pulsed electromagnetic fields(PEMFs)at 15 Hz and 1 mT 8 h/d for 3 days.A semi-quantitative RT-PCR technique was used to measure the type I collagen mRNA expression;ELISA and immunohistochemitistry techniques were used to measure type I collagen expression.Inhibitors and promoters of the cAMP-dependent protein kinase A(cAMP-PKA)pathway were added.After the cAMP-PKA pathway had been inhibited or promoted,the effects of the PEMF on type I collagen expression were measured again using ELISA and immunohistoehemistry.Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type I collagen(P≤0.01)in comparison with the controls. The type I collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added,and raised when the promoter 8-Br-cAMP was added.Conclusion PEMFs at 15 Hz 1 mT can promote type I collagen expression of bone marrow mesenchymal stem cells.and the effect is correlated with the cAMP-PKA pathway.

Objective To establish the pyrogen removing effect of activated carbon in the technics of traditional Chinese medicine (TCM) injections. Methods The content of bacterial endotoxin concentration was detected by kinetic turdimetric assay to evaluate the effect of removing pyrogen before and after using activated carbon in concentrated solution of TCM injection. Results The activated carbon adsorption rate of Shuanghuanglian concentrated solution≥70%and the activated carbon adsorption rate of Danshen concentrated solution>95%. Conclusion Pyrogen manufacturing process is scientific and rational by adding activated carbon adsorption in powder injection of TCM. The bacterial endotoxin of large doses can't be removed fully by activated carbon adsorption.