Objective To establish a HPLC method to determind the content of chlorogenic acid in Jinyin Granules. Method The mobile phase was 0.3%HAc-Methonal(74∶26). The UV detection wavelength was 327 nm. The mobile speed was 0.9 mL/min. Separation column was Kroasil ODS(4.6 mm?250 mm). Result A good linearity was obtained in the range of 65～325 ?g,regression A=24 239 869C-31 292,r=0.999 3. The average recovery was 100.10%,RSD was 0.87%(n=6). Conclusion The method was good for determining chlorogenic acid in Jinyin Granules.

Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.

BACKGROUND: Studies have shown that Gingko biloba leaf extract (GbE) is effective in promoting the functions recovery of the brain that follows traumatic injury, in improving the dysfunctions of learning and memory of the elderly, and it is also effective in improving the plasticity in central nervous system (CNS). However, what is the effect on learning and memory functions of rats with type 2 diabetes mellitus?OBJECTIVE: To study the effects of GbE on the learning and memory dysfunction and glial fibrillary acidic protein (GFAP) expressions in hippocampus of diabetic rats.DESIGN: Complete-random design, controlled experimental study.SETTING: Department of Pharmacology, Guangxi Medical University.MATERIALS: A total of 84 male Wistar rats (180-220 g), 8 weeks old,SPF, were used in this study. GbE (containing 24.8% flavone glycosides and 6.2% diterpene lactone) was purchased from Guilin Xintejia Natural plants Pharmaceutical Factory, Guangxi Province, Lot No. 200405.METHODS: The experiment was completed at the Pharmacology Lab (Provincial Lab) of the Experimental Center of Guangxi Medical University from June 2004 to March 2005. ① A total of 70 rats were rendered diabetic by a single intraperitoneal injection of streptozotocin at a dose of 55 rmg/kg dissolved in citrate buffer (pH4.4) after 24 hours fasting. Tail vein blood glucose concentration was determined 4 days later using ONE TOUCH glucose meter. A total of 56 streptozotocin-treated rats with a blood glucose concentration of ＞ 15 mmol/L were recognized as type 2 diabetic rats. ② These diabetic rats were randomly divided into model group, insulin group, high-dose GbE group, and low-dose GbE group. There were 14 rats in each group. There was no difference in the blood glucose concentration among the groups. Another 14 male rats with an intraperitoneal injection of citrate buffer solution were served as control group. After division, drugs were given. Insulin 10 μ/kg was injected subcutaneously every day for 6 months. GbE 100 mg/kg and GbE 50 mg/kg were administered through intra-gastric method every day for 6 months.The diabetic group and control group were administered normal solution through intra-gastric method every day for 6 months. ③ Six months later,Morris water maze was operated on each group of rats. The Morris water maze consisted of a large circular pool [100 cmdimension, 60 cm height,filled to a depth of 42 cm with water at (25±1) ℃]. Within the pool a submerged platform (round, black, 8 cm diameter, 2 cm below the water surface) was hidden on a fixed location, 20 cm from the edge of the pool,in which milk powder was dissolved to obscure the platform. The rat could climb on the platform to escape from the necessity of swimming. The rats were trained to locate the hidden platform. The animals were received 4 trials (2 in the morning, and 2 in the afternoon) per day on 4 consecutive days. The rat was given a maximum of 90 s to find the hidden platform.On the 5th day, the rat's learning ability was examined by observing the time to find the hidden platform (escape latency) in 90 s and the platformfinding strategy (prompt and straight way, marked 4 scores; hesitating first and then straight way, marked 3 scores; random way, marked 2 scores;aimless way and around the pool border, marked 1 score). On the 8th day,the escape latency and the platform-finding strategy were observed to examine the rat's memory level. ④ After the Morris water maze test, 8 rats of each group were sacrificed by decapitation for RT-PCR of GFAP mRNA expression, and 6 rats of each group were sacrificed for immunohistochemistry of GFAP protein expression. GFAP mRNA expression level was analyzed by the expression ratio of the interest GFAP to the control β-actin according to the computer image analysis. The GFAP protein expression was analyzed by the volume density of GFAP in hippocampus. ⑤ Data were analyzed with one-way ANOVA and q-test.MAIN OUTCOME MESURES: The effects of GbE-on the performances of the water maze Morris of type 2 diabetic rats and both GFAP mRNA and protein expressions in hippocampus.RESULTS: A total of 14 streptozotocin-treated rats with a blood glucose concentration of ＜ 15 mmol/L were rejected from the study. ① The performance of diabetic rats in the Morris water maze was significantly impaired compared to control group, the results on the 5th day and the 8th day showed that both escape latency and platform-finding strategy scores were decreased (P ＜ 0.01). The escape latency of both insulin treatment and GbE treatments on the 5th day and the 8th day was shorter than that of diabetic group, the platform-finding strategy scores was higher than that of diabetic group (P ＜ 0.05-0.01). There was no marked difference among the insulin treatment and GbE treatments groups in performance of the water maze Morris (P ＞ 0.05). ② The levels of both GFAP mRNA and protein expressions in hippocampus: Statistical analyses indicated that the level of GFAP mRNA expression of diabetic rats was significantly higher than that of the 3 other groups (P ＜ 0.01). Compared to control group, the diabetic rats showed a high immunoreactivity, the GFAP body was enlarged markedly, apophysis was obviously longer, the expressed numbers were increased, and the volume density of GFAP was also increased significantly (P ＜ 0.01). Compared to the diabetic group, the insulin treatment and GbE treatments groups showed a low immunoreactivity, the GFAP body was markedly smaller, apophysis was obviously shorter, the expressed numbers were decreased, and the volume density of GFAP was also decreased significantly (P ＜ 0.01). There were no marked differences in both GFAP mRNA and protein expressions among the insulin treatment and GbE treatments groups (P ＞ 0.05).CONCLUSION: There is cognition impairment in type 2 diabetic rats, the responsive GFAP may take a part in the progress of the learning and memory dysfunction. GbE can decrease markedly the reactive hypertrophy of astrocytes of diabetic hippocampus and improve the learning and memory dysfunction in diabetic rats.

Objective　To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods　Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results　Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions　Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.

Objective To research the method and effect about the bronchoscopic lung volume reduction by ?-cyanoac- rylate in the therapy of chronic obstructive pulmonary disease (COPD).Methods The 14 patients had been examined bosoms by CT before the operation and determined the type of emphysema and the distributing of pneumatocele,had blood gas analyzed and pulmonary function checked.The operation was carried through trachea cannula and intravenous anesthe- sia.When the bronchoscope came to the goal bronchia,we infused the meglumine diatrizoate through the biopsy orifice and approved the location of pneumatocele forward。Then,we infused erythromycin and ?-cyanoacrylate in turn through the biopsy orifice by silica del tube.Results The 3 pneumothorax patients had been removed the drainage tube in 3 days af- ter the operation.8 cases had been counterchecked sternite in one week and the pneumatocele was just like before,among which,1 case developed exudation.1 case had shown pleural thickening in the right-up lung counterchecked sternite 9 months later.1 case been checked the pulmonary function,the FEV_1 enhanced from 24.7% pred before operation to 32. 9% pred after operation one week.3 cases felt polypnea improved greatly and 7 cases felt polypnea improved a little.Con- clusion The bronchoscopic lung volume reduction by ?-cyanoacrylate is a safe,effective and economical method in the therapy of COPD.

Animal medicines mainly contain protein which was organic molecule with quaternary structure and had the property of thermal denaturation. When suffering from heat for a consistent time, the native conformation of protein would be destroyed. After denaturation the biological activity of protein will lose and some physicochemical and biochemical properties will be changed. Leech was a classical animal medicine in the views of traditional Chinese medicine (TCM) which had the functions of breaking stagnant and eliminating blood stasis. In the usage history, it was processed for a long time. No matter stir-frying leech with talc powder embodied in Chinese Pharmacopoeia or stir-baking with wine as a distinctive method in Beijing district, the process procedure was basically performed under high temperature. The purposes and intentions of process are mostly limited to technology conditions at specific historical period. In this article, based on existing processing procedure and its character of Leech, the changes of active components and pharmacological activities before and after processing under high temperature were summarized. The results demonstrate that the protein of leech would be denaturated; some active peptide such as hirudin were partly or totally destroyed; some toxic mineral elements, such as Pb, Hg, Cd, were decreased; at the same time, heating can promote some chemical components transforming into hypoxanthine which had the function of antihypertensive, antiasthmatic and antalgic. Consequently, after processed under high temperature, the purpose of decreasing toxicity and alleviating the strong property was achieved. Pharmacological changes of leech processed under high temperature were mainly manifested in the anticoagulant and antithrombotic activity, etc. Based on current processing research status about animal medicine leech, future research methods and directions on scientific connotation of leech processed under high temperature were put forward in this article.
Animals ; Biological Factors ; chemistry ; isolation & purification ; pharmacology ; Chemistry, Pharmaceutical ; methods ; Hot Temperature ; Humans ; Leeches ; chemistry

This study was aimed to observe the clinical effect of Pingai Mixture intervention on HIV/AIDS patients in order to clarify the characteristics of Pingai Mixture for AIDS treatment. A total of 41 HIV/AIDS patients who met the inclusion criteria were selected. Pingai Mixture was applied in the treatment. Three months was one treatment course. The treatment was given for two courses. Clinical symptoms, physical signs and changes in CD4+ lymphocytes of HIV/AIDS patients before and after treatment were observed. The SPSS software was used for statistical analysis. The results showed that the posttreatment total score (primary, secondary symptoms and the main physical sign cred-its) was significantly reduced compared with pretreatment (P < 0.01). The posttreatment spontaneous perspiration, night sweats, hair loss and other symptoms of a separate score was significantly reduced compared with pretreatment (P< 0.05). There were no significant changes on other symptoms or physical sign indicators. There was no significant difference in the Karnovsky score after 6-month treatment compared with pretreatment. It was concluded that Pingai Mixture is effective in AIDS treatment. It can improve the clinical symptoms and the administration is safe.

Objective: To investigate the effect of combining acupuncture and auricular point sticking on heart rate variability (HRV) in patients with post-stroke depression (PSD). Methods: A total of 80 cases with PSD were randomized into a treatment group and a control group. The control group was intervened by oral administration of paroxetine hydrochloride, whereas the treatment group received acupuncture plus auricular point sticking base on the same oral administration. The Hamilton depression rating scale (HAMD) and HRV were measured before and after treatment in both groups. Results: The individual and global scores of HAMD significantly dropped after 8 weeks of treatment in both groups (all P<0.05). In the treatment group, anxiety/somatization factor, sleep disturbance, hopelessness factor, cognition factor and global score were significantly different from those in the control group (all P<0.05). The 24 h standard deviation of all normal-to-normal R-R interval (SDNN), standard deviation of 5-minute average of normal R-R intervals (SDANN), root mean square of successive differences (RMSSD), percent of differences between adjacent normal R-R intervals >50 ms (PNN50) and high frequency (HF) were increased while low frequency (LF) and LF/HF decreased significantly after 8 weeks of treatment in both groups (P<0.05). All items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Combining acupuncture and auricular point sticking can enhance the conventional medical treatment for HRV in patients with PSD.

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

Adult ; Chordoma ; pathology ; Female ; Humans ; Nose Neoplasms ; pathology