Objective To perform a spatial analysis of myocardium uhrastructure and the activity of mitochondrial succinate dehydrogenase in sub-acute Keshan disease. Methods Myocardium samples were collected from the cases with sub-acute Keshan disease and non-myocarditis(control), and their ultrastructure was observed under electron microscope. The density of mitochondrion volume and cristal membrane and its volume were measured by a point-counting method, while mitochondrion volume was estimated by water displacement method, succinate dehydrogenase activity of mitochondrion by iron-copper method in sub-acute Keshan disease and non-myocarditis cases. Results The volume ratio of mitochondrion to the cell on myoeardium [(47.79±6.20)%], the area ratio of mitochondrion to sarcoplasm [(55.06±6.50) %], mitochondrion to myofibrils [(1.43±0,41)%], mitochondrion section area[(0.78±0.15)μm2], and ratio of the lesion of cristal membrane area to the matrix area and mitochondrion volume[(67.14±13.96)%, (44.62±13.44)%]in sub-acute Keshan disease group were obviously higher than those in control [(33.20±7.62)%, (38.07±9.43)%, (0.71±0.33)%, (0.44±0.07)μm2, (14.11± 12.51)%, (9.34±11.28)%; t = 3.75,7.93,6.61,36.40,52.65,37.51, all P < 0.05]. The volume ratio of myofibrils to cell[(34.52±5.12)%]and the area ratio of cristae mitochondria to matrix[(32.43±14.42)%]in sub-acute Keshan disease group was obviously less than those in control [(48.51±4.30)%, (86.04±12.37)%; t = 9.85, 53.46, both P < 0.05)]. Succinate dehydrogenase activity was negative in sub-acute Keshan disease group. Conclusions Myocardium ultrastructure changes in sub-acute Keshan disease including the increase of volume and areas of mitochondria and the damage of the cristal membrane in mitochondria. Succinate dehydrogenase activity is decreased or even disappeared.

BACKGROUNDMyocardial apoptosis is involved in the pathogenesis of sepsis-related myocardial depression. However, the underlying mechanism remains unknown. This study investigated the role of mitochondrial damage and mitochondria-induced oxidative stress during cardiac apoptosis in septic rats.

METHODSSeventy-two Sprague-Dawley rats were randomly divided into a control group and septic group receiving lipopolysaccharide injection. Heart tissue was removed and changes in cardiac morphology were observed by light microscopy and scanning electron microscopy. In situ apoptosis was examined using terminal transferase-mediated dUTP nick end-labeling assay and nuclear factor-kappa B activation in myocardium by Western blotting to estimate myocardial apoptosis. Appearance of mitochondrial cristae and activation of cytochrome C oxidase were used to evaluate mitochondrial damage. Oxidative stress was assessed by mitochondrial lipid and protein oxidation, and antioxidant defense was assessed by mitochondrial superoxide dismutase and glutathione peroxidase activity.

RESULTSSepsis-induced inflammatory cell infiltration, myocardium degeneration and dropsy were time-dependent. Expanded capillaries were observed in the hearts of infected rats 24 hours post-challenge. Compared with sham-treated rats, the percentage of cell apoptosis increased in a time-dependent manner in hearts from septic rats at 6 hours, 12 hours and 24 hours post-injection (P < 0.05). The expression of nuclear factor-kappa B p65 decreased gradually in the cytosol and increased in the nucleus during sepsis, indicating that septic challenge provoked the progressive activation of nuclear factor-kappa B. Mitochondrial cristae and activation of cytochrome C oxidase increased in a time-dependent manner. Both superoxide dismutase and glutathione peroxidase activities decreased, while mitochondrial lipid and protein oxidation increased between 6 and 24 hours after lipopolysaccharide challenge.

CONCLUSIONSSeptic challenge induced myocardial apoptosis and mitochondrial damage. Furthermore, mitochondrial damage via alteration of defenses against reactive oxygen species might play an important role in myocardial apoptosis during sepsis.

Animals ; Apoptosis ; physiology ; Male ; Mitochondria, Heart ; metabolism ; pathology ; Myocardium ; metabolism ; pathology ; Oxidative Stress ; physiology ; Rats ; Rats, Sprague-Dawley ; Sepsis ; metabolism ; physiopathology

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

Animals ; Apoptosis ; drug effects ; Cell Line ; Cell Proliferation ; Curcuma ; Extracellular Matrix ; metabolism ; Hepatic Stellate Cells ; drug effects ; pathology ; Plant Extracts ; pharmacology ; Plant Oils ; pharmacology ; Rats

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Cell Proliferation ; Curcuma ; chemistry ; Fas Ligand Protein ; genetics ; metabolism ; Flow Cytometry ; Hepatic Stellate Cells ; drug effects ; metabolism ; Immunohistochemistry ; Liver Cirrhosis ; pathology ; Plant Extracts ; administration & dosage ; pharmacology ; Plant Oils ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; fas Receptor ; genetics ; metabolism

AIM To investigate the effects of stir-frying,processing with butter and carbonizing by stir-frying on oil components in Gleditsiae sinensis Fructus and Gleditsiae Fructus Abnormalis.METHODS The volatile oils and fatty oils were extracted by steam distillation method and Soxhlet extraction method,respectively,after which the extraction rates were determined.GC-MS was applied to analyzing the kinds and relative contents of oil components,after which cluster analysis was performed.RESULTS After the processing,the two medicinal materials demonstrated increased extraction rates of fatty oils and decreased extraction rates of volatile oils(except for processing with butter),the extraction rates of oil components in Gleditsiae sinensis Fructus were higher than those in Gleditsiae Fructus Abnormalis,and the reduced relative contents of toxic olefin benzene components were observable.CONCLUSION The kinds and relative contents of oil components in Gleditsiae sinensis Fructus and Gleditsiae Fructus Abnormalis exist obvious differences,the former displays better medicinal quality,whose processing mechanism in alleviating dryness and strength may contribute to the reduction of relative contents of toxic olefin benzene components.

OBJECTIVETo analyze the causes and explore the measures of prevention and treatment of the cerebrospinal leak after lumbar revision surgery.

METHODSThe clinical data of 24 patients(17.78%) with cerebrospinal leak among 135 cases after lumbar revision surgery from January 2011 to January 2016 was retrospectively studied. Of them, 12 cases due to severe adhesion caused by scar formation; 4 cases due to yellow ligament hyperplasia adhesion with dura mater occurred dural tears when separating adhesion; 2 cases with severe hyperplasia and ossification of lumbar posterior longitudinal ligament occurred dural tears when revealing intervertebral disk; 2 cases due to improper nailing happened dural tears during operation. And the other 4 cases without obvious dural tears in surgery, occurred cerebrospinal leak one to two days after surgery. And the 24 patients were treated with the measures of prevention and treatment preoperatively, intraoperatively, and postoperatively.

RESULTSTwenty-four patients with cerebrospinal leak were cured after treatment and were follow-up for 6 to 30 months. No recurrence of cerebrospinal leake or local and systemic complications were found.

CONCLUSIONSScar formation is the main cause of cerebrospinal leak in lumbar revision surgery. As for lumbar revision surgery, as long as the standard control measures are taken, it can significantly reduce the incidence of cerebrospinal leak, achieve better clinical efficacy, and fundamentally solve the cerebrospinal leak problem that has plagued both doctors and patients for a long time.

Objective To find out the arsenic pollution in Yangzonghai lake and its influence to the surrounding drinking water source and the local food and to provide a scientific basis for arsenic pollution control,drinking water and food safety. Methods Arsenic monitoring were carried out in 10 points of Yangzonghai lake,45 water sources within 5 km of Yangzonghai lake, and locally produced food. Results From September 2008 to December 2009, the water arsenic concentrations of Yangzonghai lake ranged 0.06 - 0.18 mg/L, the average arsenic concentrations were 0.12 mg/L and 0.10 mg/L for 2008 and 2009, respectively, the value of 2009 was higher, and the difference was statistically significant(t = 3.284, P ＜ 0.05). There were no significant difference in the average of arsenic concentration of Yangzonghai lake between the dry and rainy season in 2009(t = 0.905, P ＞0.05). The arsenic concentrations from the No. 3 spring water ranged from 0.12 - 30.36 mg/L. After a peak value early in October 2008, the No. 3 spring water showed a downward trend month by month. In 2008 the average arsenic concentration was 23.92 mg/L, 2009 was 2.41 mg/L, down 89.92% over the previous year, the difference was statistically significant(t = 9.582, P ＜ 0.05 ). In January and February 2009, the average arsenic concentration (11.96 mg/L) was significantly higher than those of March-December(0.50 mg/L, t = 57.759, P＜ 0.05). In Yangzong town, the arsenic concentration in the drinking water and river met the national drinking water health standards. Among 78 samples monitored of the locally produced food in Yangzong town, 77 passed, with a pass rate of 98.72%. Conclusions Arsenic contamination has been effectively controlled. The drinking water and local food production has not been contaminated by arsenic except aquatic products in Yangzonghai Lake. It is recommended to take effective measures to prevent water contamination.

OBJECTIVETo investigate targeted degradation of the androgen receptor (AR) by chimeric molecules (DHT-PROTAC) via the ubiquitin-proteasome pathway in androgen-independent prostate cancer CA-2B cells, and explore the proliferation, secretion and apoptosis of the treated cells.

METHODSC4-2B cells were treated with DHT-PROTAC, and then the expressions of the AR protein and caspase3 in the C4-2B cells were detected by immunohistochemistry and Western blot. The concentration of PSA in the supernatant was examined by ELISA. The cells were counted and their proliferation analyzed by a growth curve. The inhibitory effect on the growth of C4-2B cells was evaluated by MIT assay.

RESULTSCompared with the control group, the DHT-PROTAC-treated group showed an obviously decreased expression of AR proteins with a significant attenuation of the band signals (P < 0.05), a 40% reduction of the AR-positive cells and a 60% decrease of the PSA concentration in the supernatant (P < 0.05). DHT-PROTAC exhibited an inhibitory effect on the C4-2B cells in a time-dependant manner (P < 0.05).

CONCLUSIONThe chimeric molecule (DHT-PROTAC) can target the degradation of androgen receptors, reduce the secretion of PSA and repress the in vitro growth of C4-2B cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Male ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms ; drug therapy ; metabolism ; pathology ; Receptors, Androgen ; metabolism

ABSTRACT:OBJECTIVE To observe the clinical effect and safety of treatment of Gushibao pill on severe kidney circulation deficiency and blood stasis type knee osteoarthritis.METHODS 70 patients with severe knee osteoarthritis were selected for this study,and were divided into the treatment group(38 cases)and the control group(32 cases).Patients of two groups were given health education and basic knee stick therapy treatment.On the basis of basic treatment,patients in the control group were given glucosamine hydrochloride capsules orally and treatment group were give Gubaoshi pill.WOMAC Osteoarthritis In-dex Scale and ISOA knee osteoarthritis severity index scale were used to evaluate the outcomes of two groups.Chinese medi-cine symptom score were used to evaluate the clinical symptoms.ESR,calcitonin(CT)and bone alkaline phosphatase(BALP) were detected before and after treatment.RESULTS Erythrocyte sedimentation rate in the control group significantly de-creased after a course of treatment(P <0.05),and bone alkaline phosphatase and calcitonin showed no significant change(P >0.05).Erythrocyte sedimentation rate decreased significantly in treatment group after treatment(P <0.05),and bone alkaline phosphatase and calcitonin showed no significant change(P >0.05).Scored of symptoms and signs significantly decreased in two groups,and the decreased extent of treatment group was significantly greater than that of the control group(P <0.05). Pain score,WOMAC score and ISOA scores were significantly lower in two groups of patients compared with the condition be-fore treatment(P <0.05).After treatment,pain score,WOMAC score and ISOA scores of treatment group were lower than those of the control group(P <0.05).The total effective rate of treatment group was 97.37%,and was significantly higher than the control group(80.00%).The difference was statistically significant(P <0.05).Two patients had no obvious adverse reactions during treatment.CONCLUSION Gushibao pill can decrease the level of erythrocyte sedimentation rate in patients with severe kidney circulation deficiency and blood stasis knee osteoarthritis.It is effective and safe.