AIM: To establish the method of determining procyanidin B2 content in grape seed extract by HPLC. METHODS: The determination was carried out with ZORBAX SB-C_ 18 column. The mobile phase consisted of acetonitrile-2% acetate buffer under the gradient elution condition, detection wavelength was at 280nm. RESULTS: There was a good linear relationship between the peak area and concentration in the range of 1~30 ?g/mL for procyanidin B2. The average of procyanidin B2 (n=5) was 99.29% and RSD=1.64%, respectively. CONCLUSION: The method is simple, accurate, reproducible and can be used for assay of procyanidin B2.

Objective:To observe dynamic change of phosphorylated tau protein and non phosphorylated tau protein in axon and explore whether GSK3 in serum were related with phosphorylated tau protein in brain of EAE mice.Methods: Mice were randomly divided into six groups:EAE group of acute stage,EAE group of paralytic stage,EAE group of remission stage,control group of acute stage,control group of paralytic stage,control group of remission stage,each group had twelve mice.EAE model were constructed by MOG35-55 peptides in EAE group, the saline was used in control group.The nerve function scores and incidence were observed and compared.We observed degree of brain inflammation by HE staining and measured axons which contained two kinds of tau protein by immunohistochemistry.GSK3 in serum was tested to find correlation with phosphorylated tau protein in brain by ELISA method.Results:Nerve function scores in EAE group were higher than control group.The degree of inflammation damage was more serious in EAE group than control group,gradually enhancing with time.Phosphorylated tau protein in brain gradually increased before mice paralyzing(P<0.01),but it decreased when symptom relieved(P<0.01).GSK3 in serum were correlated with phosphorylated tau protein in brain(r=0.9326,P<0.01),linear regression equation:Y=2.950+0.418X.Conclusion:Phosphorylated tau protein in brain are correlated with axon damage and GSK3 in serum could indirectly reflect axon damage in brain.

AIM: To develop a rapid HPLC method for quality control of traditional Chinese medicinal ingredients consisted of citrus flavonoids, naringin, hesperidin, neohesperidin, sinensetin and nobiletin. METHODS:Gradient elution with non-salt mobile phase ( methanol and water only) HPLC method on a Kromasil column ( 100-5C18-250A, 4.6 mm ×250 mm, 5 μm, C18 reverse phase) with peaks identification through DAD full UV wavelength scan. UV 284 nm and 332 nm profiles were observed. RESULTS: Satisfactory resolution, linearity, 95%～ 102% of recovery and 1.88 ～ 2.93% of repeatability were obtained for those five citrus flavonoids. Content of 6 Citrus aurantium L. based TCM ingredients were analyzed and identified. CONCLUSION: Rapid HPLC test method on citrus flavonoids was developed and can be in LC-MS identification.

BACKGROUND: Cleistocalyx operculatus is a dried alsbastrum of myrtle. It is reported that cleistocalyx operculatus extracts can improve cardiac contraction through inhibiting the activity of Na+/K+-ATPase, and decrease rate of contraction. Do cleistocalyx operculatus extracts have the biological activity of antioxidation?OBJECTIVE: To observe the effects of cleistocalyx operculatus on oxidative injury of PC12 nerve cells induced by H2O2.DESIGN: Non-randomized controlled study.SETTING: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology.MATERIALS: The experiment was conducted at New World Institute of Biotechnology, State Key Laboratory of Bioreactor Engineering of East China University of Science and Technology, from May to November 2002.Eight adult male Kunming mice were selected. PC12 nerve cells were supplied by Shanghai Cell Institute of the Chinese Academy of Sciences.METHODS: Model of oxygenic injury of PC12 nerve cells was estabPC12 cells were cultured in 96-well plates. Cleistocalyx operculatus was diluted with RPMI1640 culture medium into five concentrations of 0.001,0.01, 0.1, 0.5 and 1 g/L with 3 wells in each concentration; each well had 2×103 cells. Blank control group, or non-drug culture medium group, was set. Under the standard condition, cells were cultured for 48 hours and ascells were inoculated in 96-well plate with the density of 2×103 for 24-hour wall adhering, and then divided into normal control group (normal cell without H2O2 or cleistocalyx operculatus extracts), 0, 0.01, 0.1, 0.5 and 1 g/L cleistocalyx operculatus. Cells in all groups except normal control group were treated with 200 μmol/L H2O2 at 37℃ for 3 hours, then cleistocalyx operculatus of various concentrations was added and survival rate was asfree radicals: PC12 cells with oxygen-derived free radicals were treated in the same way as done for cell survival rate assay and measured with CDCFH staining method.fect of cleistocalyx operculatus extracts on intracellular and extracellular oxygen-derived free radicals in PC12 nerve cells induced by oxidative injury.operculatus could protect nerve cells; however, at 0.055-1.00 g/L the effect on cell growth did not significantly differ from that of blank control extracts had no protective effect on the injury of PC12 nerve cells induced by H2O2. At 1.00 g/L, it had strong plerosis for oxidative injury of PC12 and extracellular oxygen-derived free radicals in PC12 nerve cells was increased; however, at 0.01 g/L concentration of cleistocalyx operculatus extracts, the level was lower than that in model group.dation of membrane lipid of hepatic microsome, but also protect against oxcleistocalyx operculatus extracts is related to its concentration. At 1.00 g/L,it has great capacity of oxidation plerosis, and at 0.01 g/L it can decrease the level of oxygen-derived free radicals inside and outside cells.

Objective To characterize the skin physiology function of patients with ache and to facilitate its treatment. Methods Sixty patients with acne (20 males and 40 females) and 60 healthy human controls (20 males and 40 females) were included into this study. The average age of patients and controls was 23.4 years and 25.1 years, respectively. Sebumeter was used to detect the sebum secretion in the following areas: forehead, nose, right and left cheeks, Cutometer(R) MPA580 to measure the skin elasticity, and Scalar Moisture Checker to test the skin hydration on right and left cheeks. Results A significant increase was observed in the level of sebum secretion in the T-zones (199.98±58.21 μg/cm2 vs 117.55±63.16 μg/cm2, t=7.34, P<0.05) as well as in the cheeks(154.45±55.06 μg/cm2 vs 87.50±47.36 μg/cm2, t=7.14, P< 0.05) in the patients compared with the controls. However, the level of skin elasticity and hydration was of no significant difference between the patients and controls (0.7931±0.0755R vs 0.7882±0.0498R, 30.75%±3.87% vs 30.94%±2.91%, respectively, both P>0.05). Conclusion Facial sebum secretion is increased in patients with acne.

BACKGROUND: The expression of main histocompatibility antigen in cornea of rats is similar to that of human. It is verified in early experiments that penetrating corneal transplantation in rats is more repetition and reliability, and the operation difficulty is lower than that in mice.OBJECTIVE: To establish rat model of penetrating corneal transplantation and analyze the reasons of complications during model preparation and the corresponding managements and probe into the method for improving the success rate of rat model of corneal transplantation.DESIGN: Randomized controlled experiment was designed.SETTING: Department of Ophthalmology , Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Department of Ophthalmology in Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from March to May 2004. Seventysix outbred female Wistar rats of two months old were employed and randomized into experimental group and control group with 38 rats in each,without eye disease, of clean grade, body mass varied from 180 to 200 g and their right eyes were taken as acceptors. Thirty-eight SD female rats were employed and their both eyes were taken as donors. All of the animals were provided from Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology.METHODS: Central penetrating corneal transplantation in situ was adopted and the model was established on the right eye of Wistar rat. Ten minutes before operation, tropicamide eye drops was administrated twice to amplify adequately the pupil of the right eye up to 4.0 mm. According to body mass, general anesthesia was done with abdominal injection of 3.0 mL/kg chloral hydrateand dicaine eye drops was applied twice 5 minutes before the operation with 0.1 volume fraction. Routine sterilization was done on the head of rat, the eyes kept in horizontal level and respiration kept smooth. The clip in dental-ophthalmology department was used to fix the posterior of the operated eye and make it to be semiluxated. The ring driller 3.5 mm in diameter was used to collect corneas of both eyes in SD rat as the implant tissue and place in culture dish with endothelial part upward, covered with methyl cellulose of 20 g/L mass weight concentration. The ring driller, 3.0 mm in diameter was used to collect the cornea of the right eye in Wistar rat to be the graft foreman, and the implant tissue was sutured into the graft foreman on the right eye of Wistar rat with 10-0nylon thread intermittently. Eight stitches were done in corneal transplantation in the experimental group and 6 stitches in the control group. After operation, ofloxacin eye drops was administrated and the operated eye was covered with plaster. Corneal rejection was observed and the evaluation was done in 3 items, named, turbidity, edema and neogenetic vessel. The total score of 3 items was taken as the rejection index on the day. Rejection was determined if the rejection index on the day ≥5 or corneal turbidity reached the 3nd grade. x2 and Fisher definite probability methods were used for statistical examination.plant tissue and corneal rejection.RESULTS: Of 76 Wistar rat acceptors, 5 rats were dropped out, among which, 1 rat was died accidentally after operation in the experimental group, 2 rats were died accidentally after operation in the control and 2 rats were injured cornea on the operated eyes due to fighting. But by supcomplications after corneal transplantation: In the experimental group, the number with anterior adhesion of sclera, non-round pupil; limited wound healing and non-formation of anterior chamber was lower than that in the implant tissue and corneal rejection: the survival time of corneal implant tissue was similar basically in both experimental group and the control [(8.9±2.3), (8.6±2.3) days, P ＞ 0.05]. Rejection presented in both groups in 16 days after corneal transplantation and the incidence of rejection was 100%.CONCLUSION: With stitches maintained and without intervention, the acute rejection is similar in rat corneal transplantation and human corneal transplantation. It is explained in the experiment that 8 stitches in corneal transplantation in which the rat implant tissue of 3.5 mm matches graft foreman of 3.0 mm can reduce the complications of rat corneal transplantation and becomes a satisfactory animal model of corneal transplantation.Skillful microscopic operation techniques, satisfactory operation instruments and adequate amplilfied pupil can reduce to the most extent the postoperation complications in rats.

Objective To evaluate the application of Addenbrooke′s cognitive examination revised(ACE?R)scale to evaluate cognitive function in patients with chronic kidney disease(CKD). Methods We used ACE?R scale to evaluate the cognitive function on 205 cases of CKD patients received outpatient and inpatient treatment in renal Department of four hospitals in Nanjing. One?way ANOVA analysis ,receiver operating characteristic (ROC)curve and other statistical methods were adopted to compare differences. Results (1)There is significant difference of ACE?R scale scores among group NC?NCDs(89.18 ± 4.80),group Mild?NCDs(77.28 ± 5.80)and group Major?NCDs(55.90 ± 10.90)(F = 292.28,P < 0.01). There is significant difference between any two groups(all P<0.01).(2)(ACE?R)scale(AUC=0.944,P<0.01)showed significantly higher sensitivity than mini?mental state examination(MMSE)scale(AUC = 0.777,P < 0.01)on identifying Mild?NCDs patients with chronic kidney disease(P < 0.01);(3)The optimal cut?off value of total ACE?R score between group NC?NCDs and group Mild?NCDs was 83/84,(sensitivity 88.00%,specificity 85.90%,Youden index 0.739),the optimal cut?off value between group Mild?NCDs and group Major?NCDs was 83/84(sensitivity 89.70%,specificity 96.10%, Youden index 0.858). Conclusion ACE?R possesses a good ability in distinguishing different severity of cognitive impairment in patients with CKD. ACE?R can effectively identify early mild cognitive impairment ,which is suitable for clinical application as a tool to evaluating and studying of cognitive function in patients with CKD.

Objective To evaluate the effect of group cooperative learning on the experimental teaching of obstetrics and gynecology nursing.Methods We chose 113 undergraduate nursing students,using the traditional self-learning mode in the experimental teaching of cleaning and disinfection of the vulva and the group cooperative learning mode in the experimental teaching of vaginal douching.Their practical manipulations of the nursing skills by the two teaching modes were compared.A self-designed questionnaire was adopted to investigate the student's attitude to the group cooperative learning.Results The examination achievement by the group cooperative learning was better than that by the traditional self-learning,(Z=-4.986,P<0.001).89.4%of the students approved of the group cooperative learning.Conclusion The group cooperative learning may improve the effects of experimental training,contribute to the cooperative spirits of students,and also make full use of the human resources in teaching.

This article was aimed to study the mechanism of interaction between human serum albumin (HSA) and zinc ions, in order to provide the information on the secondary structure modification of HSA and the thermodynamics parameters using circular dichroism (CD) and isothermal titration calorimetry (ITC). CD and ITC were applied in the study. The concentration of HSA were 0.025 mmol·L-1, 0.05 mmol·L-1, 0.1 mmol·L-1, and 0.2 mmol·L-1, respectively. The concentration of zinc ion was 5 mmol·L-1. The CD analysis revealed that the secondary structure modification of HSA differed depending on the concentration of HAS. And the content ofα-helix was inversely proportional to the concentration of HSA. When the concentration of HSA was 0.2 mmol·L-1, the content ofα-helix was special. A series of thermodynamics parameters including association constants (Kb), stoichiometry (N), entropy (ΔS) and enthalpy (ΔH) were investigated by ITC analysis. And two types of binding sites were observed when ZnSO4(mmol·L-1)?HSA (mmol·L-1)= 5?0.2. The secondary structure modification of HSA interacting with zinc ions depended on the concentration of HSA, a dramatic reduction ofα-helix was detected when the concentration of HSA attained 0.2 mmol·L-1. And the protein hydrophobicity reduction and peptide chains expansion were equally observed at this concentration. The ITC analysis also revealed endothermic and exothermic binding sites in the ZnSO4-HSA interaction when ZnSO4 (mmol·L-1)?HSA (mmol·L-1)= 5?0.2, indicating that the endothermic sites were specific but not preferential for zinc ions interactions. These results provided theoretical supports for the application of Zn2+ to open the endothermic sites of HSA.

Objective To establish the optimal extracted method for content dete rmination of naringin in Citrus grandis. Methods RP-HPLC was used to determinat e the content of naringin extracted with the above two methods from different ye ar samples of Citrus grandis. Results The average content extracted with ultraso nic extraction was 13.53 %,and the average content extracted with soxhlet extr action was 11.98 %,there being insignificant difference between the two method s. Conclusion The content of naringin extracted with ultrasonic extraction is mo re than that with the soxhlet extraction,which be receipted in Chinese pharmeco pia. And ultrasonic extraction method is more convenient and can save time.