Objective To investigate the prognostic factors of life and functional outcome of the first hemiparetic stroke patients. Methods One hundred and eighteen stroke subjects were registered prospectively. The Barthel index (BI) , Rankin scale (RS) , Mortricity index(MI) , Mini-mental state examination (MMSE) , Montgomery-Asberg depression scale (MADRS) and a scale of general state and risk factors were used to evaluate at the 48th hour, the 15th day and the 90th day after stroke. Results The patients' performance, as demonstrated by their scores with all the evaluation instruments, changed significantly at all the time points of evaluation after stroke (P ＜0.05). There was no significant difference between the performance at the 48th hour and the 15th day after stroke ( P ＞ 0.05 ). But at the 90th day after stroke, the activity of daily living performance and the depression status recovered significantly (P ＜ 0.01 ). Logistic regression analysis showed that, such factors as pneumonia, urinary incontinence within 48th hour and deep sensation disturbance might adversely influence patients' activity of daily living performance at the 90th day after stroke; the muscle strength of upper extremities at the 48th hour, and MMSE scores at the 15th day after stroke acted as the protective factors. Conclusions The stroke patients improved significantly with regard to their clinical and functional manifestations when evaluated 90 days after stroke onset. The main factors influencing the activity of daily living performance 90 days after stroke onset included deep sensation disturbance,pneumonia, urinary incontinence and muscles strength of upper extremities at 48th hour, and MMSE scores at the 15th days after onset.

Objective To study the effects of helicase-like transcription factor (HLTF)transfection on DNA repair protein level in radiation-induced apoptotic cells.Methods Human lung carcinoma A549 cells were cultured and transfected with FLAG-tagged wild type HLTF (wild type HLTF transfection group),RING structure domain (ubiquitin conjugating region) mutatation HLTF expressing plasmid (mutant transfection group),empty plasmid (congtrol group) respectively.And the other cells were used as mock transfection group.All cells were irradiated with 15 Gy of 60Co γ-rays to induce apoptosis.Western blotting was used to detect the protein levels of the DNA repair proteins HRAD17 and HRAD52 in the transfected cells.Results The levels of HRAD17 and HRAD52 in the wild type HLTF transfection group was significantly lower than that of the control group.There was no significant difference in HRAD17 and HRAD52 levels between the mock transfection group and ubiquity in conjugating region mutation group.complexes of HLTF and HRAD17 and HRAD52 could be found in the irradiation-induced cells.Conclusions HLTF mediates the degradation of HRAD17 and HRAD52 in the irradiation-induced apoptotic cells possibly by the interaction of the protein complex causing ubiquitination of the repair proteins.

OBJECTIVE:To optimize the extraction technology of Bushen huoxue capsules. METHODS:Central composite de-sign-response surface method was used to optimize the extraction technology with the amount of water and boiling time as main fac-tors using normalized value of the contents of icariin and salvianolic acid B,the yield of dry extract as index. Validation test was al-so conducted. RESULTS:The optimal extraction technology was as follows as 15-fold water,reflux extracting for 75 min,extract-ing for 2 times. The deviation of measured value and predicted value was 0.000 3% in validation test. CONCLUSIONS:The cen-tral composite design-response surface method can be used to optimize the extraction technology of Bushen huoxue capsules with good prediction,and the technology is simple and stable.

Objective To compare the effect of enteral nutrition by jejunum colostomy nutrition infusion pump of patients after Whipple surgery as well as reduce adverse reactions in patients.Methods Sixty-five cases with the implementation of Whipple and jejunum of colostomy were selected as our subjects,who were hospitalized in the Affiliated hospital of Hebei United University from Feb.2009 to Nov.2013.All patients were divided into observation group (33 cases) and control group (32 cases) according to the methods of nutrient input.Patients in observation group were given nutrition infusion pump pumping (15 to 50 ml/h) ;and patients in control group were adopted disposable infusion connection infusion with the speed of 30 drops/min with the thermostat heating temperature and the water pipe.The blood glucose,serum albumin,blood electrolyte concentration of postoperative,and the adverse reactions during input nutrient solution including vomiting,abdominal distention,diarrhea and other adverse circumstance were recorded.Results At 1st,3rd,5th day,there was no statistically significant difference in terms of the levels of glucose,blood albumin,blood C1,Na +,K + between two groups(blood glucose:F inner grouP =3.01,P ＞ 0.05 ; F between group =2.90,P ＞ 0.05 ; F cross group =2.87,P ＞ 0.05 ; serum albumin:F inner group =2.94,P ＞ 0.05 ; F between group =2.89,P ＞ 0.05 ; F cross group =2.76,P ＞ 0.05 ; blood Cl:F inner group =1.78,P ＞ 0.05 ; F between group =1.96,P ＞ 0.05 ; F cross group =1.88,P ＞ 0.05 ; blood Na +:F inner group =1.06,P ＞ 0.05 ; F between group =1.35,P ＞ 0.05 ; F cross group =1.27,P ＞ 0.05 ; blood K +:F inner group =3.12,P ＞ 0.05 ; F between group =3.04,P ＞ 0.05 ; F cross group =2.93,P ＞ 0.05).There were significant differences regarding of the rate of vomiting,abdominal distention,diarrhea and other adverse conditions compared with the infusion enteral nutrition has good clinical effect,postoperative blood (x2 =4.029,4.381,4.905 respectively; P ＜ 0.05).Conclusion The methods of colostomy enteral nutrition with infusion pump after Whipple surgery is proved to be with the better clinical effect in reducing postoperative vomiting,abdominal distention,diarrhea and other adverse conditions compared with the infusion enteral nutrition,and there are no significant difference in the terms of the levels of glucose,blood albumin,blood Cl,Na +,K +.

OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits

Objective To evaluate preventive effect of polysaccharides from Lycium ruthenicum Murray against photodamage in HaCaT cells,and to explore its possible mechanism.Methods Ultrasoundassisted extraction was used to extract polysaccharides from Lycium ruthenicum Murray in Qaidam Basin.In vitro cultured HaCaT cells were randomly divided into 3 groups:control group receiving no treatment,ultraviolet B (UVB) group irradiated with 30 nJ/cm2 UVB alone for 1 hour,experimental group pretreated with 2 g/L Lyeium ruthenicum polysaccharide solution followed 6 hours later by 30 mJ/cm2 UVB radiation for 1 hour.At twelve hours after UVB radiation,an inverted microscope was used to observe cell morphology.Then,MTS assay was performed to estimate cell proliferation,flow cytometry to detect cell apoptosis,an enzyme-labeled antigen method to detect levels of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px),as well as to evaluate the activity of superoxide dismutase (SOD) and catalase (CAT),and enzyme-linked immunosorbent assay (ELISA) to measure levels of intedeukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in HaCaT cells and their culture supernatant.Results Compared with the control group,the UVB group showed obscure cell morphology,cell death and floating phenomenon,while cells became swollen but renained morphologically distinct in the experimental group.MTS assay revealed that the cell proliferative activity significantly differed among the above 3 groups (F =48.88,P ＜ 0.01),and the cell proliferative activity was significantly lower in the UVB group (1.72 ± 0.12) than in the control group (2.34 ± 0.11,P ＜ 0.05) and experimental group (2.11 ± 0.10,P ＜ 0.05).Moreover,the apoptosis rate was significantly higher in the UVB group (82.41％ ± 2.49％) than in the control group (3.98％ ± 0.19％,P ＜ 0.05) and experimental group (22.79％ ± 0.97％,P ＜ 0.05),as well as higher in the experimental group than in the control group (P ＜ 0.05).Compared with the control group,the UVB group showed significantly higher levels of MDA,supernatant levels of IL-1 and TNF-α,and intracellular levels of TNF-α,but significantly lower GSH-Px levels and activity of SOl and CAT (all P ＜ 0.05).However,there was no signifi-cant difference in the intracellular level of TNF-α between the UVB group and control group (P ＞ 0.05).Conclusion Lycium ruthenicum polysaccharide has protective effects against photodamage in HaCaT cells,likely by reducing the synthesis and secretion of inflammatory substances as well as free radicals.

The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.
Caffeic Acids ; analysis ; toxicity ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; toxicity ; Quinic Acid ; analogs & derivatives ; analysis ; toxicity ; Xanthium ; chemistry ; classification

To develop different methods for determining siRNA content and the entrapment efficiency of siRNA loaded liposomes, SYBR Gold electrophoresis method and Ribogreen fluorospectrophotometry method were used respectively. SYBR Gold electrophoresis method has a good linear relation in a range at 0.2-2.0 micromol x L(-1) (R = 0.9930), and the recovery at the high, middle and low concentrations were 96.35%, 96.92%, and 100.74%, respectively (n = 3). The intra-day and inter-day RSD were far below 5% (n = 5). Ribogreen fluorospectrophotometry method has a good linear relation in a range at 10-50 nmol x L(-1) (R = 0.9971), and the recovery at the high, middle and low concentrations were 98.22%, 99.88% and 99.64%, respectively (n = 3). The intra-day and inter-day RSD were far below 5% (n = 5). The content and the entrapment efficiency of three batches of siRNA cationic liposomes were 98.52%, 97.85% and 99.20%, 96.45%, respectively, with these two methods. And there is no significant difference by ANOVA. Both of the two methods are accurate, sensitive, convenient method for determination of the siRNA content and the entrapment efficiency of siRNA loaded cationic liposomes.
Drug Carriers ; Drug Delivery Systems ; Electrophoresis ; Liposomes ; chemistry ; RNA, Small Interfering ; analysis ; Spectrometry, Fluorescence

Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.
Angelica ; chemistry ; Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cytarabine ; administration & dosage ; Humans ; K562 Cells ; Leukemia ; drug therapy ; Liver ; drug effects ; Male ; Mice ; Mice, SCID ; Polysaccharides ; administration & dosage

In this study, OVA-induced asthma mice was taken as the model, and orally administered with different concentration of ethanol extracts of crude and processed Stemona tuberosa, in order to determine the cytokine level released from Th1 and Th2 in splenocytes. RT-PCR was carried out to determine the genetic expression of T-bet/GATA-3 in lung, and compare the differentiation between ethanol extracts of crude and processed S. tuberosa in therapeutic effect on asthma in mice. According to the results, compared with the crude samples, processed samples significantly increased the levels of inflammatory factor INF-gamma (P < 0.05) and decreased IL-5 (P < 0.05) in splenocytes. According to the RT-PCR results, the administration of processed samples could increase the ratio of T-bet/GATA-3 (P < 0.05). The experiment showed that ethanol extracts of both crude and processed S. tuberosa could treat asthma by regulating Th1/Th2 ratio, but processed samples showed more notable effect. This indicated that crude and processed S. tuberosa had significant pharmacological difference. Therefore, it was more rational to apply processed S. tuberosa in clinical treatment of asthma and chronic cough, which layed a foundation for further revealing the processing mechanism of S. tuberosa.
Administration, Oral ; Animals ; Asthma ; drug therapy ; immunology ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; GATA3 Transcription Factor ; metabolism ; Gene Expression Regulation ; drug effects ; Mice ; Mice, Inbred BALB C ; Stemonaceae ; chemistry ; T-Box Domain Proteins ; metabolism ; Th1 Cells ; drug effects ; secretion ; Th2 Cells ; drug effects ; secretion