Humans ; Purpura, Thrombocytopenic ; diagnosis ; therapy

OBJECTIVE: To observe the effect of acupuncture on activities of microglia in traumatic brain injury (TBI) rats. METHODS: Fifty-four male SD rats were randomly and equally divided into normal control, model and acupuncture groups according to the random number table (n＝18 rats in each group). The TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone (to induce the left parietal cerebral contusion). Acupoints "Baihui" (GV20), "Shuigou" (GV26), "Fengfu" (GV16), "Yamen" (GV15) and bilateral "Hegu" (LII4) were stimulated intensively by twirling the filiform needles with force at a range of >360° and a frequency of 160－180 cycles/min for 10 sec in every acupoint, once every 5 min during the 15 minutes' needle retaining. The treatment was given once every day for successive 14 days. The rats of the normal and model groups were grabbed and fixed with the same procedure. The behavioral changes were tested using modified neurological severity score (mNSS). The histopathological changes of the injured cerebral cortex tissues were observed by using hematoxylin-eosin (H.E.) staining, and the fluorescence intensity of Iba-1 (marker of microglia) positive products in the surrounding tissue of the cerebral focus was displayed by immunofluorescence staining, and the contents of neuron specific enolate (NSE) and neurite outgrowth inhibitor-A (Nogo-A) in serum (indicating a secondary nerve damage) were assayed by ELISA. RESULTS: The mNSS scores were significantly increased on day 1, 3, 7 and 14 in the model group in comparison with the normal group (P<0.01) and considerably decreased at the 4 time-points after acupuncture intervention relevant to the model group (P<0.05, P<0.01). H．E. staining showed that modeling induced pathological changes such as the excursion of cell nucleus, cellular swel-ling, vacuole-like change, neuron death, karyopyknosis dissolution, and proliferation of fibrous tissue were relatively milder in the acupuncture group. The average fluorescence intensity values of Iba-1-positive products, serum NSE and Nogo-A contents on day 3, 7 and 14 were significantly higher in the model group than in the normal group (P<0.05, P<0.01), and notably down-regulated in the acupuncture group than in the model group (P<0.05, P<0.01, except Nogo-A on day 3). CONCLUSION: Acupuncture intervention may accelerate neurological function recovery in TBI rats, which is closely related to its effects in inhibiting the activation of microglia and secondary nerve damage.

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.

Administration, Inhalation ; Adrenergic beta-Agonists ; adverse effects ; Child ; Extremities ; Humans ; Male ; Paralysis ; chemically induced ; Terbutaline ; administration & dosage ; adverse effects

Adult ; Aged ; Alopecia ; chemically induced ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Bone Neoplasms ; drug therapy ; pathology ; secondary ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; secondary ; Cisplatin ; adverse effects ; therapeutic use ; Humans ; Leukopenia ; chemically induced ; Liver Neoplasms ; drug therapy ; pathology ; secondary ; Lung Neoplasms ; drug therapy ; pathology ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Organoplatinum Compounds ; administration & dosage ; Paclitaxel ; administration & dosage ; Remission Induction ; Survival Rate ; Taxoids ; adverse effects ; therapeutic use ; Young Adult

The role of methyl-CpG binding domain protein 2 (MBD2) in an ApoE-deficient mouse model of age-related macular degeneration (AMD) was investigated. Eight-week-old Mbd2/ApoE double deficient (Mbd2(-/-) ApoE(-/-)) mice (n=12, 24 eyes, experimental group) and MBD2 (wt) ApoE(-/-) mice (n=12, 24 eyes, control group) were fed on Western-type diet for 4 months. The mice were sacrificed, and total serum cholesterol levels were analyzed and Bruch's membrane (BM) of the eyes was removed for ultrastructural observation by transmission electron microscopy. Moreover, intercellular adhesion molecule 1 (ICAM-1) immunoreactivities were evaluated by fluorescence microscopy in sections of the eyes in both groups for further understanding the function mechanism of MBD2. There was no significant difference in the total serum cholesterol levels between control group and experimental group (P>0.05). Transmission electron microscopy revealed that AMD-like lesions, various vacuoles accumulated on BM, notable outer collagenous layer deposits and dilated basal infoldings of retinal pigment epithelium (RPE) were seen in both groups, and the BM in control group was significantly thickened as compared with experimental group (P<0.05). Fluorescence micrographs exhibited the expression of ICAM-1 in choroid was higher in control group than in experimental group. We are led to conclude that MBD2 gene knockout may lead to accumulation of more deposits on the BM and influence the pathogenesis of AMD via triggering endothelial activation and inflammatory response in choroid, improving microcirculation, and reducing lipid deposition so as to inhibit the development of AMD-like lesions. Our study helps to provide a new therapeutic approach for the clinical treatment of AMD.

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.

This article introduces an overview of management system for conflict of interest of the European Pharmacopoeia Commission (EPC) and the United States Pharmacopoeia Convention (USP). The EPC and USP have standardized the management system for conflict of interest in drug standard work in multiple management documents, such as the Guide for the Work, Code of Practice for the Work, Form for Declaration of Interests and Confidentiality Undertaking of the EPC, bylaws, Rules and Procedures of the Council of Experts, Code of Ethics, Standards of Conduct of the USP, in order to ensure the transparency and fairness of drug standard development, improve the credibility and rigor of drug standards. This article introduces the management system for conflict of interest of the EPC and USP, providing reference for the improvement of relevant management systems of the Chinese Pharmacopoeia Commission.

ObjectiveTo characterize the neural progenitor cell in the human amnion mesenchyme and epithelial layer with specific mark proteins of neural stem cell.MethodsExpressions of specific mark proteins of neural stem cell including nestin, glial fibrillary acidic protein (GFAP), musashi-1, vimentin and PSA-NCAM in human amnion tissue and cultured amniotic cells were determined by immunohistochemistry and immunofluorescence staining.ResultsExpressions of pluripotent neural stem cell specific makers (nestin, musashi-1, vimentin and PSA-NCAM) were detected in the human amnion mesenchyme and epithelial layer. In addition, cultured amniotic cells were expressed several neural stem cell specific markers including nestin, GFAP and PSA-NCAM. Nestin+ and GFAP+ double positive cells were identified in the human amnion tissue and cultured amniotic cells by immunohistochemistry and immunofluorescence staining.ConclusionSpecific mark proteins of neural stem cell are expressed in human amnion tissue and cultured amniotic cells.

AIM To establish a RP HPLC method for the determination of zidovudine(AZT) plasma concentration. METHODS A shim pack,VP ODS. 5 ?m 4 6?150 mm radial compression column was used with a mobile phase of tetrahydrofuran water trifluoroacetic acid(10∶89 8∶0 2). The flow rate was 1 0 ml?min -1 .The detector was operated at 267 nm. RESULTS The calibration curves of AZT were linear separately within the concentration range of 0 02～20 mg?L -1 . The relative standard deviations for within day and between days assays were all less than 5%. The relative recovery of the method were 96 4%. The limit of quantitation of the assay is 10 ?g?L -1 of plasma. CONCLUSIONS The methed is rapid,accurate and stable. The determination of AZT plasma concendration could be used.