Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

Objective To explore the risk factors in elderly type 2 diabetes mellitus with cerebral infarction. Methods Retrospective investigarion was performed on 148 elderly hospitalized patients with type 2 diabetes.The patients were classified based on the presence or absence of cerebral infarction and compared with 60 controls.Logis- tic regression analysis was used to reveal the risk factors for cerebral infarction.Results The levels of systolic blood pressure(SBP),body mass index (BMI),fasting blood glucose (FBG),total cholesterol (TC),triglyceride (TG) and plasma fibrinogen(Fg) were higher in the patients with cerebral infarction[141.15?17.46)mmHg,(23.81?3.53)kg/m~2,(8.82?2.81)mmol/L,(5.69?1.15)mmol/L,(2.08?0.75)mmol/L and (4.08?0.65)g/L] than those without cerebral infarction[(129.78?14.65) mmHg,(22.18?3.22)kg/m~2,(7.06?1.72 )mmol/L,(5.09?1.12)mmol/L,(1.62?0.43)mmol/L and (3.48?0.58)g/L].The logistic analysis showed COUR,SBP,FBG, TC,TG and Fg were the independent risk factors for cerebral infarction.Conclusion Early intervention of the inde- pendent risk factors including SBP,FBG,TC,TG and Fg in elderly patients with type 2 diabetes was important for reduction and postponement of cerebral infarction.

OBJECTIVETo observe the differential effect of joint ultrasound on the syndrome differentiation of rheumatoid arthritis (RA) by observing the high frequency ultrasound performances among inactive stage and different syndromes in active stage.

METHODSTotally 83 RA patients in the active stage were assigned to the dampness heat syndrome group (DHS, 59 cases)and the cold dampness syndrome group (CDS, 24 cases) according to Chinese medicine (CM) syndrome typing. Besides, 20 RA patients in the remission stage were recruited as the control group (abbreviated as the remission group). By using high frequency ultrasound and power Doppler ultrasound technology, a comparative observation of synovitis, tenosynovitis, synovial blood flow, and bone erosion in the 2nd-5th metacarpophalangeal (MCP) joints, proximal interphalangeal (PIP) joints, wrist joints, knee joints, the second and the fifth metatarsophalangeal (MTP) joints (a total of 24 joints) was performed in all patients. Correlation analyses were performed between the ultrasound performance, laboratory indices, and the disease activity. Ultrasound data of each RA patient were analyzed by their total scores. Χ2 test was used for enumeration data. The measurement data was expressed as x ± s. One-way ANOVA was used for data of normal distribution, while non- parametric test was used for data of non-normal distribution. Correlation analysis of two variables was performed for clinical indicators and ultrasound indicators. Its significance was detected using Pearson correlation.

RESULTSCompared with the remission group, the severity degree of synovitis, tenosynovitis, synovial blood flow, and bone erosion significantly increased in the DHS group (P < 0.01). There was statistical difference in ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF (P < 0.05, P < 0.01). There was statistical difference in the severity degree of synovitis and synovial blood flow, and DAS28 score in the CDS group (P < 0.05). Compared with the CDS group, there was statistical difference in the four ultrasound indices (P < 0.05, P < 0.01), ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF in the DHS group (P < 0.05, P < 0.01). There was no statistical difference in G, IgG, IgA, or IgM among the three groups (P > 0.05). There existed positive correlation between ESR and the synovitis degree, synovial blood flow, and bone erosion in the DHS group (r = 0.444, 0.397, 0.486, P < 0.05).There existed positive correlation between ESR and the synovitis degree, bone erosion, and synovial blood flow in the DHS group (r = 0.378, 0.270, P < 0.05). There existed positive correlation between the DAS28 score and the synovitis degree and synovial blood flow in the DHS group (r = 0.304, 0.351, P < 0.05).

CONCLUSIONSThe inflammation degree was the most severe in RA patients of DHS. High frequency ultrasound could provide better evidence for Chinese medical syndrome differentiation of RA patients.

Arthritis, Rheumatoid ; diagnostic imaging ; Humans ; Medicine, Chinese Traditional ; Metacarpophalangeal Joint ; ultrastructure ; Syndrome ; Synovitis ; diagnostic imaging ; Ultrasonography

OBJECTIVETo subjectively and objectively assess the effect of Jiawei Xiaoyao Powder (JXYP) on sleep in patients with psychological stress insomnia. METHHODS: A randomized controlled study was conducted in 33 patients with psychological stress insomnia. They were assigned to 4 groups, 4 in the TCM group treated with JXYP, 5 in the Western medicine (WM) group treated with Estazolam, 9 in the integrated medicine (IM) group treated with JXYP plus Estazolam, and 10 in the control group treated with placebo. Quality of sleep in patients was assessed subjectively and objectively before treatment and 6 weeks after treatment by Pittsburgh sleep quality index (PSQI), self-rating scale of sleep (SRSS) and polysomnography (PSG), respectively.

RESULTSSubjective assessment on sleep showed that after 6-week treatment, the scores of PSQI and SRSS remarkably reduced in the TCM, IM and control groups (P < 0.05), while the decrease was insignificant in the WM group (P > 0.05), but no significant difference between groups was shown. The objective assessment by PSG showed that no significant change was found after treatment in parameters of total sleep time (TST), sleep time of phase 1 and 2, slow wave phase, rapid-eye-movement (REM) phase, sleep latency, REM sleep latency, also in long waking and short waking times in all group (P > 0.05), but a significant increase of sleep efficacy (P < 0.05) and an increasing trend of TST (P > 0.05) were shown in the IM group, and an increasing trend of both in the TCM group (P > 0.05).

CONCLUSIONJXYP, combined with or without Estazolam, can improve the quality of sleep subjectively, and the combination of the two could enhance the efficacy of sleep in patients with psychological stress insomnia.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Estazolam ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Sleep Initiation and Maintenance Disorders ; drug therapy ; psychology ; Stress, Psychological ; complications ; Young Adult

In this study, hydrophilic matrix sustained release tablets of total lactones from Andrographis paniculata were prepared and the in vitro release behavior were also evaluated. The optimal prescription was achieved by studying the main factor of the type and amount of hydroxypropyl methylcellulose (HPMC) using single factor test and evaluating through cumulative release of three lactones. No burst drug release from the obtained matrix tablets was observed. Drug release sustained to 14 h. The release mechanism of three lactones from A. paniculata was accessed by zero-order, first-order, Higuchi and Peppas equation. The release behavior of total lactones from A. paniculata was better agreed with Higuchi model and the drug release from the tablets was controlled by degradation of the matrix. The preparation of hydrophilic matrix sustained release tablets of total lactones from A. paniculata with good performance of drug release was simple.
Andrographis ; chemistry ; Delayed-Action Preparations ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydrophobic and Hydrophilic Interactions ; Kinetics ; Lactones ; chemistry ; Tablets ; chemistry

OBJECTIVETo study the effectiveness of sequential occlusal adjustment for kinetic food impaction.

METHODS36 patients who claiming food impaction with normal proximal contact were examined and analyzed about their occlusal relationship and configuration. Sequential occlusal adjustment was made to reduce pestle-mortar-like cusp, to create food escaping groove and to decrease mesial surface of the distal tooth cusp. One week, two weeks and six months later, the patients were reexamined and their oral conditions were evaluated.

RESULTSAn elimination of food impaction was claimed for 32 patients in one week appointment and all 36 patients in two-week appointment. Six months later, no patient reported food impaction.

CONCLUSIONThe use of sequential occlusal adjustment results in an effective elimination of some kind of kinetic food impaction.

Food ; Humans ; Mastication ; Occlusal Adjustment ; Tooth

OBJECTIVETo probe into the best acupoint-prescription for the simple acupoint catgut embedding therapy in treatment of bronchial asthma.

METHODSForty-eight Wistar rats were randomly divided into six groups: the simple acupoint catgut embedding at Feishu (BL 13), Danzhong (CV 17) and Shenshu (BL 23) group (group A), the simple acupoint catgut embedding at Shenshu (BL 23) group (group B), the simple acupoint catgut embedding at Feishu (BL 13) and Danzhong (CV 17) group (group C), Dexamethasone group (group D), model group (group E), and control group (group F), 8 rats in each group. The asthmatic models were established by Ovalbumin (OVA) except group F. Rats in group A, B and C were treated with catgut embedding at the corresponding acupoints from the first experimental day. In group E and D, Dexamethasone and sterile were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistantly. No interventions were added on group F. For the six groups, the symptoms of asthmatic attack were observed and the pathologic changes of lung tissue were examined.

RESULTS(1) The times of sneeze and rhinocnesmus scratching nose in group E were increased significantly compared with those in group F (both P < 0.01), but there was no significant difference between group D and F (both P > 0.05). As compared with that in group E, except the times of sneeze in group C, the times mentioned above were decreased significantly in all the treatment groups (all P < 0.01). There was no significant difference between group A and D (both P > 0.05), but the times mentioned above were increased significantly in group B and C as compared with that in group A (all P < 0.01). (2) The symptoms of asthmatic attack and the pathologic changes of airway tissue were all alleviated in group A, B and C, but a better amelioration was observed in group A, with no mucus epistom in the bronchial lumen and few infiltrations of inflammatory cells around the bronchi.

CONCLUSIONThe improvement of the simple acupoint catgut embedding at "Feishu" (BL 13), "Danzhong" (CV 17)and "Shenshu" (BL 23) on the airway inflammation in asthmatic rats is better than that of catgut embedding at "Feishu" (BL 13) and "Danzhong" (CV 17) or at "Shenshu" (BL 23) only.

Acupuncture Points ; Acupuncture Therapy ; methods ; Animals ; Asthma ; pathology ; therapy ; Catgut ; Male ; Medicine, Chinese Traditional ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo study the clinical and pathological features of children with Duchenne muscular dystrophy (DMD), with the aim of increasing the possibility of early diagnosis.

METHODSThe clinical data of 50 children who were definitely diagnosed with DMD, based on clinical manifestations and the results of skeletal muscle biopsies and monoclonal antibody immunohistochemical staining, was reviewed.

RESULTSThe children showed similar clinical manifestations, including running slowly in the toddler period, muscle weakness when climbing stairs and standing up followed by squatting down and walking abnormalities a predominant increase in serum creatine kinase level increased dominantly, and myopathic lesions seen on electromyography. Hematoxylin-eosin staining showed similar pathological presentations in all 50 children, including different-sized muscle fibers with rounding, degeneration and necrosis in various degrees, and proliferation of connective tissues. There was some inflammatory cell infiltration in muscle fibers and interstitial tissues. Dystrophin expression was completely absent at the sarcolemma in all 50 children, and sarcoglycan-α,-β, -',-δ expression was reduced to various degrees in 33 of them.

CONCLUSIONSFor children with the clinical manifestations mentioned above, skeletal muscle biopsies and monoclonal antibody immunohistochemical staining are recommended as these examinations contribute to a definite diagnosis of DMD by demonstrating dystrophin deficiency at the sarcolemma.

Adolescent ; Child ; Child, Preschool ; Dystrophin ; genetics ; Humans ; Immunohistochemistry ; Infant ; Male ; Muscle, Skeletal ; pathology ; Muscular Dystrophy, Duchenne ; complications ; genetics ; pathology ; Retrospective Studies

OBJECTIVETo determine whether cysteinyl leukotriene receptor agonist LTD(4) and cysteinyl leukotriene receptor 1 (CysLT(1)) antagonist pranlukast affect the differentiation of human neuroblastoma SK-N-SH cells.

METHODSSK-N-SH cell morphological changes induced by LTD(4), pranlukast and LTD(4) + pranlukast were observed with retinoid acid (RA) as the positive control. The expressions of CysLT(1) and CysLT(2) receptors were detected by immunoblotting analysis, and the expression of microtubule-associated protein-2 (MAP-2), a neuron marker, was detected by fluorescent immunostaining.

RESULTThe immunoblotting results showed that SK-N-SH cells expressed CysLT(1) receptor moderately, and CysLT(2) receptor highly. The morphological results showed that RA, pranlukast and LTD(4) + pranlukast induced the compaction of the cell bodies and the outgrowth of neurites, while LTD(4) had no significant effect. The immunostaining results showed that MAP-2 was distributed in the cell bodies in control or pranlukast-treated cells; it was distributed in cell bodies and neuritis in RA-treated cells. Pranlukast increased the numbers of MAP-2-positive cells.

CONCLUSIONThe CysLT(1)receptor antagonist pranlukast modulates the differentiation of SK-N-SH cells.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Chromones ; pharmacology ; Humans ; Immunoblotting ; Immunohistochemistry ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Neuroblastoma ; metabolism ; pathology ; Receptors, Leukotriene ; metabolism

OBJECTIVETo investigate let-7c's effect on the proliferation of human hepatocellular carcinoma cell HCCLM3 by transient transfection and the mechanism inside.

METHODSLipofectamine 2000 was used to transfect miRNAs into HCCLM3 cells. The cells were divided into three groups, let-7c group: let-7c was transfected, negative control group: negative control miRNA was transfected, blank control group: nothing was transfected. The proliferation of HCCLM3 cells was evaluated using Cell Counting Kit-8 (CCK-8). The cell cycles of each group were assayed by flow cytometry. Western blot and Real time PCR were used to analyze the protein and mRNA expressions of cyclin D1. Statistical analysis was performed with SPSS 17.0.

RESULTSThe absorbances of let-7c group were 0.70 ± 0.05, 0.77 ± 0.09 at 48 h and 72 h after transfection, lower than that of blank control group (0.97 ± 0.10, 1.21 ± 0.12) and negative control group (0.91 ± 0.07, 1.12 ± 0.09), 48 h: F = 14.431, P < 0.05, 72 h: F = 21.146, P < 0.05. The flow cytometry at 72 h after transfection revealed that let-7c increased the percentage of cells in G1 phase. The percentage of blank control group was 43.53% ± 0.86%, the negative control group was 44.82% ± 0.77%, and the let-7c group was 54.52% ± 0.13%, F = 240.739, P < 0.05. let-7c suppressed expressions of cyclin D1 at both protein and mRNA levels. The protein levels of cyclin D1 were 0.48 ± 0.09, 0.47 ± 0.06 and 0.23 ± 0.06 (F = 11.316, P < 0.05) in blank control group, negative control group and let-7c group, respectively. The mRNA levels were 1.03% ± 0.29%, 1.01% ± 0.11% and 0.63% ± 0.14% (F=6.315, P < 0.05) in the above three groups, respectively.

CONCLUSIONLet-7c can inhibit proliferation of HCCLM3 cells and increase the proportion of cells in G1 phase. The mechanism may be that let-7c represses the expressions of cyclin D1 at both protein and mRNA levels.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Humans ; Liver Neoplasms ; genetics ; pathology ; MicroRNAs ; genetics ; RNA, Small Interfering ; Transfection