Objective To investigate current situation of prescription of traditional Chinese medicine (TCM) for acute stroke patients.Methods From June 2008 to October 2010,with simple cluster sampling,48 centers were randomly assigned to the intervention group receiving a standardized treatment strategy,and the usual care group without specific recommendations for treatment.No specific recommendations or restrictions of TCM were proposed for both groups.Consecutive hospitalized patients with acute ischemic stroke were recruited in this study.Clinical data and treatment strategy (including TCM) were recorded by trained investigators.The prescription of TCM three months prior to admission and at discharge was recorded and analyzed.The chi-square test was used for analysis of qualitative data.Results Totally 3 664 patients were included in the analysis (1 709 patients in standard care group and 1 955 patients in the usual care group) with male to female ratio 2∶ 1.The difference of distribution of age,sex and education level between the two groups was not statistically significant.One hundred and eighty-seven patients (5.10％) had taken TCM three months prior to admission,81.28％ of them only taking one type of TCM,without significant difference between the two groups.The percentage of patients taking TCM at discharge was significantly increased to 30.65％ for all the patients with higher percentage (38.16％) in the usual care group than that in the standard care group (22.06％).The percentage of using more than one type of TCM was also higher in the usual care group (x2 =141.43,P ＜ 0.01).Totally,112 types of TCM were prescribed at discharge.Conclusions Nearly one third of hospitalized patients with acute stroke were prescribed TCM at discharge,with mostly monotherapy and a great variety.

Objective Lipid metabolism disorder can lead to male sterility, but its mechanism of affecting spermatogenesis and testis microenvironment remains unclear.This study aimed to investigate the effects of high fat diet on reproductive function and the expression of Ox-LDL within testis in male mice. Methods 16 C57BL/6 mice aged 8 weeks were divided into two groups by random number table method.High fat diet group was fed with high fat diet while normal fat diet group was fed with normal fat diet .At the end of 16 weeks, the levels of TG, TC, Ox-LDL and testosterone in serum were measured.The sperm concentration and motility from caudal epi-didymis were analyzed.The testis structure was observed by HE stai-ning.The localization and expression of Ox-LDL in testis were detec-ted by immunohistochemical technique. Results Compared with the normal diet group mice, the body weight (P<0.05) and the ser-um levels of TC [(2.31 ±0.33)mmol/L vs (6.54 ±0.31)mmol/L, P<0.01], Ox-LDL [(0.32 ±0.03)mg/L vs ( 0.44 ±0.06)mg/L, P<0.01] increased in high fat diet group mice significantly.The serum level of TG was of no significant difference(P>0.05).The serum level of testosterone[(3.64 ±0.43)mg/L vs (0.40 ±0.14) mg/L, P<0.01], the sperm concentration[(9.95 ±0.75)106/mL vs (5.66 ±1.51)106/mL, P<0.01] and the sperm motility[(54.69 ±17.84)%vs (32.48 ±5.80)%, P<0.01] decreased in high fat diet group significantly.HE staining also showed that the amount of Leydig cells, spermatids and spermatozoons reduced obvi-ously in high fat diet group.Immunohistochemistry staining showed that Ox-LDL was mainly distributed around Leydig cells of mice tes-tis.The expression of Ox-LDL in high fat diet group increased significantly ( P<0.01) . Conclusion The expression of Ox-LDL in Leydig cells of high fat diet C57BL/6J mice increases significantly, which may inhibit testosterone biosynthesis and affect spermatogen-ic function.

AIM: To evaluate the effects of the extract of Buddleja officinalis eye drops in basic tears secretory volume, tear film stability, expression of androgen receptors(AR) in castrated rats with dry eye, and to investigate the therapeutic effects of extract of Buddleja officinalis on dry eye caused by gonadal hormones level imbalance. METHODS:A total of 45 Wistar masculinity rats were divided at random into 9 groups, including normal group(A1,A2 and A3), model group(B1,B2 and B3), therapy group with extract of Buddleja officinalis eye drops(C1,C2 and C3). The "1" stood for being fed for 1 month, and "2" for 2 months, and "3" for 3 months. The dry eye model was established with orchiectomy on group B,C. Group C was treated with Buddleja officinalis extract eye drops for one month. All rats were checked with Schirmer Ⅰ test (SⅠt) and tear film break-up time (BUT). Expression of AR was analyzed by flow cytometer(FCM). RESULTS:The SⅠt value of group C was significantly higher than that of group B (P<0.01) and the BUT value of group C was significantly longer than that of group B (P<0.01), which indicated the eye drop could significantly keep basic tears secretory volume and tear film stability. And the expression of AR of group C was much higher than that of group B,which showed that available composition of the eye drops maybe display androgen-like activity.CONCLUSION:The main components of extract of Buddleja officinalis is the flavonoids which could significantly inhibit happening of dry eye of rat after androgen level lowered. Its mechanism is like androgen's and it could display androgen-like activity to keep basic tears secretory volume and tear film stability.

For the investigation of the metabolic regulation of Saccharomyces cerevisiae under different culture conditions, the proteins of cell utilizing various carbon sources were separated by two-dimensional electrophoresis with immobilized pH gradients as the first dimension and SDS-PAGE as the second. Samples were taken in the log phase of batch culture using glucose or lactic acid as carbon source, while another sample was taken from the broth when glucose was consumed up and ethanol accumulated in the previous phase was further metabolized. After electrophoresis, the protein spots were detected by silver-stain in a Hoefer Automated Gel Stainer with a protein silver staining kit. Silver-stained gels were scanned and digitized to create computer images. About 500 protein spots were detected by employing the 2D proteome image analysis system Image Master 2D Elite and SWISS-2DPAGE proteome database. Most of the protein expressed and involved in the glycolysis, pentose phosphate (PP) pathway, anaplerotic pathway, as well as TCA cycle were analyzed. The metabolism regulation of protein level for Saccharomyces cerevisiae under various carbon sources, as well as during different phase of growth, was studied. The expression of several glycolytic enzymes (glk, pgi, pgk, eno, pyk) was up-regulated while the expression of enzymes in oxidative pentose phosphate pathway (zwf, gnd) was down-regulated when ethanol and lactic acid were taken as carbon source. Simultaneously, frucotose 1,6-biphosphatase was found to be up-regulated due to the gluconeogenic requirement. Citrate synthase and Malate dehydrogenase do not exhibit significant difference, indicating TCA cycle is necessary when utilizing glucose, ethanol or lactic acid as carbon source. Thus, the NADPH loss due to the repressed pentose phosphate pathway could be compensated by TCA cycle in cases of ethanol and lactic acid. The expression of malic enzyme and isocitrate lyase are activated to a large extent when metabolizing ethanol, indicating glyoxylate shunt is essential in transferring ethanol to generate four carbon precursors for the biosynthesis and the NADP-dependent malic enzyme could also serve as compensation mechanism for NADPH loss in this case.
Carbon ; metabolism ; Culture Media ; Culture Techniques ; methods ; Electrophoresis, Gel, Two-Dimensional ; methods ; Ethanol ; metabolism ; Lactic Acid ; metabolism ; Proteome ; analysis ; metabolism ; Proteomics ; Saccharomyces cerevisiae ; genetics ; growth & development ; metabolism ; Saccharomyces cerevisiae Proteins ; analysis ; metabolism

OBJECTIVETo investigate the protective effect and action mechanism of petroleum ether extracts from Saussurea involucrate on brain tissues of hypoxia rats under constant pressure and closed conditions.

METHODThe PESI dosage-dependent experiment for hypoxia rats was conducted under constant pressure and closed conditions by intraperitoneally injecting 125, 250, 500 mg x kg(-1) to finalize that the optimum dosage is the high dose of PESI. Afterwards, 90 Wistar rats were randomly divided into the hypoxic model group, the acetazolamide 250 mg x kg(-1) group and the PESI high dose group. Each group was further divided into three subgroups according to different hypoxia times, with 10 rats in each subgroup. Under the same hypoxia and administration conditions, the rats were sacrificed after 0, 3, 6 h respectively. Their brain samples were collected for common pathological observation and immunohistochemical staining of HIF-1alpha. Real-time RT-PCR was used to detect HIF-1alpha, EPO, HO-1 and Caspase-3 gene expressions. And the Western blot assay was adopted to detect HIF-1alpha protein expression.

RESULTThe brain tissues of the hypoxia model group were severely damaged with the increase in the hypoxia time. The acetazolamide group and the PESI high does group were damaged in a much lower degree. According to the gene expression and the Western blot assay, high dose of PESI could inhibit HIF-1alpha expression. According to the pure gene expression test, high dose of PESI could increase EPO and HO-1 mRNA expressions, but inhibit Caspase-3 mRNA expression.

CONCLUSIONPESI's protective mechanism for brain tissues of hypoxia rats under constant pressure and closed conditions may be related to its effects in inhibiting HIF-1alpha expression, increasing EPO expression and resisting cell apoptosis.

Alkanes ; chemistry ; Animals ; Brain ; cytology ; drug effects ; metabolism ; Caspase 3 ; genetics ; Cell Hypoxia ; drug effects ; Cytoprotection ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Erythropoietin ; genetics ; Gene Expression Regulation, Enzymologic ; drug effects ; Heme Oxygenase-1 ; genetics ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Male ; Rats ; Rats, Wistar ; Saussurea ; chemistry

Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

OBJECTIVEThis study investigated the 8003 base pair (bp) fragmentation damage of brain mitochondrial DNA in newborn piglets at different times after hypoxic-ischemic brain damage (HIBD) so as to explore the biomolecular foundation of neonatal neuronal metabolic disorders.

METHODSFifty 3-day-old piglets were randomly assigned into Control and HIBD groups. The HIBD group was subdivided into groups sacrificed at 0, 24, 48 and 72 hrs post-HIBD (n=10). HIBD was induced by left carotid ligation and exposure to 8% oxygen for 2 hours. The Control group was exposed to air and was sham-operated. The left hippocampal cortexes of all subjects were obtained to amplify the fragments of 200 bp and 8003 bp by the LX-PCR method. The PCR products were electrophoresed on agaros gels to obtain the integral optical density (IOD).

RESULTSThe IOD of 8003 bp fragment was markedly reduced in the HIBD 0 hr group (22.616 +/- 2.276) when compared with that of the Control group (56.995 +/- 0.317) (P < 0.05). The IOD value remained lower at 24 hrs (27.719 +/- 0.309) and 48 hrs post-HIBD (49.491 +/- 3.233) (P < 0.05). Until 72 hrs post-HIBD, the IOD (55.972 +/- 2.236) restored to the control value.

CONCLUSIONSThe brain mitochondrial DNA was fragmented in newborn piglets following brain hypoxia-ischemia. It did not recover to normal until 72 hrs post-HIBD. The fragmentation damage of mitochondrial DNA may be related to the depression of mitochondrial respiratory enzymes activity and neuron apoptosis.

Adenosine Triphosphate ; metabolism ; Animals ; Apoptosis ; DNA Damage ; DNA, Mitochondrial ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; Nitric Oxide ; physiology ; Polymerase Chain Reaction ; Swine

The src homology 2 (SH2)-domain containing inositol-5-phosphatase (SHIP) is another recently identified lipid phosphatase after phosphatase and tensin homology deleted on chromosome ten gene (PTEN). It plays an important role in negatively regulating the proliferation of hematopoietic cells. The relationship between SHIP and the inhibition of tumor proliferation is rarely reported. The purpose of this study is to evaluate the apoptosis induced by SHIP gene in K562 cell line and to explore the involved signaling pathway. The K562 cells were transfected with human SHIP gene by using the lentiviral vector containing SHIP, and the transfection was verified by fluorescent quantitative PCR (FQ-PCR) and Western blot. Then the effects of SHIP protein expression on cell growth and apoptosis were measured. The levels of p-Akt, bcl-2 family, caspase and the activity of NFkappaB were assayed by Western blot and ELISA, respectively. The results are as follows: (1) Human leukemia cell line K562 was SHIP-negative; (2) Transfection with SHIP gene led to the re-expression of SHIP mRNA and protein in K562, as shown by FQ-PCR and Western blot; (3) The expression of SHIP protein inhibited cell growth and significantly increased apoptosis in K562 cells; (4) Compared to that in control group, the expression level of p-Akt-308 and p-Akt-473 in SHIP-expressing cell group decreased significantly (P<0.01); SHIP activated caspase-9, caspase-3, up-regulated protein levels of bad, p27, down-regulated expression of bcl-xL, while it had no effect on the expression of bcl-2 and bax. Furthermore, the inhibition of NF-kappaB was achieved along with the inactivation of Akt. These data suggest that SHIP gene has potential abilities to inhibit K562 leukemic cell proliferation and induce its apoptosis via inactivating PI3K/Akt pathway. The loss of SHIP might be the explanation of aberrant high-level p-Akt in human leukemia. It may be at least one of the mechanisms by which the loss of SHIP expression contributes to leukemia progression.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Proliferation ; Down-Regulation ; Genetic Vectors ; Humans ; Inositol Polyphosphate 5-Phosphatases ; K562 Cells ; NF-kappa B ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphoric Monoester Hydrolases ; genetics ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo analyze CD21 expression in diffuse large B cell lymphoma (DLBCL) and to explore its relationship with the clinicopathological characteristics and prognosis.

METHODSThe clinical data from 80 DLBCL patients who were treated in First Hospital of Jilin University from June 2005 to September 2011 were retrospectively analyzed. The cases were subjected to immunohistochemical staining (SP method) for Ki-67, CD20, CD79a, CD3, CD43, CD5, cyclin D1, bcl-2, CD10, bcl-6, GCET-1, FOXP-1 and MUM-1 protein expression in the tumor tissue. Immunohistochemistry was also used to detect CD21 expression in the tumor tissue. SPSS 18.0 was used to analyze the relationship between CD21 expression and various clinical factors, and the relationship between various clinical factors including CD21 and overall survival.

RESULTSIn the patients aged under 60 years, the incidence of CD21(+) lymphoma (64.0%, 16/25) was significantly higher than that of CD21(-) lymphoma (38.2%, 21/55). There were more CD21(+) lymphoma patients who were at clinical stages I-II (52.0%, 13/25) than patients with CD21(-)lymphomas (23.6%, 13/55). There were also more CD21(+) lymphoma patients (68.0%, 17/25) having less than two extranodal sites involvement than CD21(-)lymphoma patients (41.8%, 23/55). In addition, there were more CD21(+) lymphoma patients with IPI 0-2 (68.0%, 17/25) than CD21(-)lymphoma patients (41.8%, 23/55). There were more CD21(+) lymphoma patients with GCB subtype (60.0%, 15/25) than CD21(-)lymphoma patients (23.6%, 13/55). Death related to DLBCL was less in CD21(+) lymphoma patients (32.0%, 8/25) than CD21(-) lymphoma patients (56.4%, 31/55). Univariate analysis showed that these clinical pathological characteristics affected the overall survival of DLBCL patients, including age, ECOG score, LDH, extranodal involvement, IPI index, CD21 expression, treatment option and efficacy (P < 0.05) . Cox multivariate analysis showed that ECOG score, LDH, extranodal involvement, CD21 expression were closely related to prognosis, and the difference was statistically significant (P < 0.05). Among the 80 patients, the overall survival (OS) of CD21(+) lymphoma patients was significantly higher than that of CD21(-) lymphoma patients.

CONCLUSIONSThe expression of CD21 is associated with young age at onset, early clinical stage, small number of involvement and low IPI index. The OS and median overall survival of CD21(+) lymphoma patients are significantly higher than those of CD21(-) patients. CD21 expression, ECOG score, LDH, extranodal involvement are independent prognostic factors in DLBCL, and in particular, the expression of CD21 is more significant in the prognosis of DLBCL patients.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Child ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Gastrointestinal Neoplasms ; pathology ; Germinal Center ; pathology ; Humans ; Immunohistochemistry ; L-Lactate Dehydrogenase ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prednisone ; therapeutic use ; Prognosis ; Receptors, Complement 3d ; metabolism ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use ; Young Adult

BACKGROUNDMiddle mediastinal masses comprise a wide variety of tumors but may also reflect lymphadenopathy, and thus remain an interesting diagnostic challenge. We performed positron emission tomography (PET) of mediastinal masses in order to evaluate the ability of PET to predict the malignancy of these tumors. We compared histologic findings, videomediastinoscopy, computed tomography (CT), and PET-CT in patients with mediastinal disease.

METHODSThirty-two patients were evaluated with CT, PET-CT and videomediastionoscopy, and all studies were performed within four weeks in each patient. (11)C-choline as a PET tracer was used to visualize masses. PET data were evaluated using the standardized uptake value (SUV) and were compared with pathologic data.

RESULTSThere were 13 men and 19 women aged from 21 to 74 (mean 45.2) years. Among the patients with mediastinal diseases, sarcoidosis was diagnosed in 12 patients, tuberculosis in 5 patients, lymphoma in 5 patients, and noncaseating granulomata without classical "sarcoid" finding in 3 patients. N2 or N3 nodal metastasis was revealed in 6 of 7 patients who had non-small cell lung cancer or suspected lung cancer, and one was negative (the pathological diagnosis was reactive hyperplasia). The accuracies for correctly diagnosing mediastinal masses for CT, PET-CT and videomediastinoscopy were 38% (12/32), 63% (20/32), and 91% (29/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 11.130, P < 0.001). The SUVs were similar among these diseases. On the other hand, if the diagnostic classification was benign vs malignancy, the accuracies for CT, PET-CT and videomediastinoscopy were 53% (17/32), 75% (24/32), 100% (32/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 22.042, P < 0.001). The SUV of malignant lesions (6.9, 3.2 - 9.8; n = 11) appeared to be higher than that of benign lesions (4.9, 2.9 - 8.3; n = 21), however, this difference was not statistically significant (P = 0.054).

CONCLUSIONSTo diagnose lesions located in the middle mediastinum, videomediastinoscopy possesses the highest diagnostic accuracy, and therefore remains the gold standard. PET-CT is valuable for differential diagnosis of benign vs malignant lesions, CT alone or PET alone (SUV) may provide misdiagnosis in a substantial proportion of patients with mediastinal masses.

Adult ; Aged ; Carbon Radioisotopes ; Female ; Humans ; Male ; Mediastinal Diseases ; diagnosis ; Mediastinoscopy ; methods ; Middle Aged ; Positron-Emission Tomography ; Tomography, X-Ray Computed ; Video Recording