Objective To detect the effect of osthole on proliferation and apoptosis of HL-60 cells and its molecular mechanism.Methods HL-60 cells proliferation was measured through the CCK8 assay method.The cell morphological changes were observed by Hoechst33342 staining after 8 h of drug effect.Induction of apoptosis was determined by flow cytometry and fluorescent microscopy.Expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR,and the expressions of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL were evaluated by using western bolt assay.Results Osthole could inhibit the proliferation of HL-60 cells,the maximum inhibiting rate was (90.7 ±4.5)％,F =138.46,P =0.000; the apoptosis rate was 33.6％,F =27.75,P =0.006.The changes of apoptosis of cells and nucleus were shown in cell morphological observation.Osthole affected the decrease of the mRNA levels of Bcl-2 and the increase of the Bax mRNA levels via a dosedependent manner(F =210.12,P =0.000).Western blotting demonstrated that osthole could lead to the increase of the expression levels of cleaved caspase-3,caspase-8,caspase-9,Fas and FasL in the HL-60 cell line via a time-dependent manner.Conclusion Data suggests that osthole inhibits proliferation and induces apoptosis of HL-60 cells through mitochondria-dependent pathway and death-receptor pathway.

Objective To analyze the effects of self-ligating bracket on enlarging the maxillary alveolar arch and to investigate the the non-extraction treatment programs.Methods All patients were used heat activated basic and precise theory for the orthodontist in order to make a plan fordeveloping nickel-titanium wire,then assessed by occlusalplane analysis method.The maxillary model andcephalometric radiographs were collected and measured before the treatment.The data of the resultswereanalyzed.Results After the treatment,the increase of the width of the dental arch was related to the /FMA,crowding degree and canines initial position.The increase of arch length was related to the ∠ANB,crowding degree,malocclusion classification,canines initial position andthe second molar eruption.Conclusion The essential factors include the crowding degree of arch,∠FMA,∠ANB,the angle of the anterior teeth,malocclusion classification,canines initial position andthe second molar eruptioncan affect the result of non-extraction quick self-ligating bracket appliance treatment on enlarging the maxillary alveolar.

Objective To analysis the clinical manifestations of mtDNA A3243G mutation in adulthood.Methods The clinical features were investigated in 36 cases (28 patients from 5 families with the mutation and 8 sporadic cases),in whom mtDNA A3243G mutation was confirmed genetically in 23 cases (15 cases from 5 mutation families and 8 sporadic cases).Cranium radiology was performed in 14 cases.Muscal biopsies were performed in l0 cases.Results Among 28 cases in the 5 family,there were 9 cases (32.1%) with stroke like episodes,17 cases (60.7%) with diabetic mellitus and 16 cases (57.1%) with deafness.Such symptoms usually combined with each other and rarely existed alone. Cardiomyopathy and renal failure were uncommon.In the 23 cases with mtDNA A3243G mutation,14 cases (61.0%) had mitochondria] myopathy,encephalopathy,lactic acidosis,and stroke-like episodes (MELAS),mostly presenting cognitive abnormalities,dysarthria or aphasia and headache,3 cases (13.0%) were asymptomatic carriers,2 cases (8.7%) had autonomic dysfunction,2 cases (8.7%) had diabetic mellitus with or without nerve deafness,1 case (4.3%) had diabetic mellitus with infertilitas and cardiomyopathy,respectively.Cranial radiological images revealed the changes more commonly in the temporal and occipital lobes and less frequently in the frontal lobes.Ragged red fibers were confirmed in 9 of 10 cases with muscle biopsies.The proportion of mutant mtDNA A3243C was not significantly different between MEALS (28.75%?13.69%) and non-MELAS (25.08%?11.54%).Conclusions mtDNA A3243G mutation mainly results in the lesions in the central nerve system,pancreatic island and acoustic nerve in adulthood.Heart and kidney are less frequently involved.Cognitive abnormalities,aphasia and headache are the major symptoms of adult MELAS.Families have with more than 1 patient with diabetic mellitus and deafness,indicating that the mutation is other than MELAS mutation.We should pay more attention to the non-MELAS symptoms in the families with mtDNA A3243G mutation.

Objective To explore the molecular types of methicillin-resistant Staphylococcus aureus (MRSA) strains present in major hospitals in Qingdao area, using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods, trying to find out the epidemiological characteristics of these MRSA isolates. Correlation of the PFGE types with microbiological phenotypes and clinical data was also studied. Methods 360 isolates of MRSA were procured during 2003 to 2007 from major hospitals in Qingdao. PFGE technology was applied to comparatively analyze the chromosomal DNA digested with endonuclease Sma Ⅰ . Comparison of DNA fragments patterns from each MRSA strain and cluster analysis were performed with the Bionumericus version ' 2.0' software. A dendogram was generated using PFGE macrorestriction fragments on gel images. Data was used to predict the possibility of each PFGE type via SPSS software version 11.0, using the variables as predictors including groups on patient's age, gender, source and the site where MRSA was isolated. Antibiotic sensitivity patterns of these MRSA isolates were determined by K-B tests, and a correlation between these patterns and PFGE types was investigated. Housekeeping genes were amplified with PCR and sequenced in representative strains of variant PFGE types to identify their allelic profile. Results 5 types of PFGE patterns (M0-M4) were identified with MI being the predominant and M2 next to it which was significantly correlated to the isolates from wounds. M3 type strains were mainly isolated from ICU wards and there were a few cases complied with M4 type with no correlated variant factors found in this study. A unique pattern of MRSA isolates with its M0 distinct from other types had not been reported. No significant association was found between PFGE individual types,gender or age groups. M1 and M2 types were the major proportional PFGE patterns among different hospitals. No vancomycin-resistant isolates were detected among 360 MRSA strains. No significant association was found between individual antibiotic resistance and specific PFGE types. Data from MLST analysis showed that the aUelic profiles of M1 and M3 type strain had the same ST239 linage which was commonly present in China. For M2 and M4 representative strains, the allelic profiles were ST5 and ST240, respectively. ST45 and ST398 were corresponding to two PFGE patterns clustered as M0 type. Conclusion Nosocomial infection due to MRSA was evenly distributed among different age groups and no gender bias was observed. The PFGE types of MRSA strains isolated in major hospitals in Qingdao were highly correlated with the sources of isolates and ST239 isolate seemed the prevalent and widespread one. Strategies should be designed to further monitor and prevent or minimize the spread of ST5 MRSA isolates and the like, in Qingdao area.

The teaching status of Molecular Pharmacognosy in 28 institutions in China was investigated by questionnaire and the survey data was analyzed by SPSS. Research contents included course beginning years, majors, class hours, characteristics of the course, teaching ways, the theory and practice contents, evaluation modes, selection of teaching material, teaching achievements, teachers and so on for undergraduates and graduates. Research results showed that with 20 years' development, Molecular Pharmacognosy had been offered for both undergraduate and graduate students in at least 20 colleges and universities and Molecular Pharmacognosy education in China showed good development momentum. At the same time, to promote the development of Molecular Pharmacognosy further, investment for it should be increased and practical teaching condition should be improved.
China ; Humans ; Molecular Biology ; education ; manpower ; methods ; trends ; Pharmacognosy ; education ; manpower ; methods ; trends ; Plants, Medicinal ; chemistry ; genetics ; Surveys and Questionnaires ; Teaching ; manpower ; methods ; trends

OBJECTIVETo explore the dynamic changes of transforming growth factor-α (TGF-α) and transforming growth factor-β1 (TGF-β1) of liver cirrhosis induced by multiple pathogenic factors in rats.

METHODSAnimals in the cirrhosis group were fed a mixture of maize flour, lard, cholesterol and alcohol plus subcutaneously injection with carbon tetrachloride (CCl₄), the CCl₄(0.5 ml/100 g · w) was injected at the first day of experiment and the 40% CCl₄oil solution (0.3 ml /100 g · w) was injected at an interval of three days. The thirty-six male SD rats were randomly divided into liver cirrhosis group of the 4th, 6th and 8 th week, and normal control group of the 4th, 6th and 8th week. The contents of alanine transferase (ALT), endotoxin, tumor necrosis factor-α (TNF-α) and homocysteine (Hcy) in plasma were evaluated. Histopathological changes of the liver were observed under microscope with the staining of HE. The expressions of TGF-α and TGF-β1 were analyzed by the method of immunohistochemistry.

RESULTSCompared with the corresponding normal control group, the levels of ALT, endotoxin, TNF-α and Hcy in plasma were gradually significantly increased in liver cirrhosis group of the 4th, 6th and 8th week (P < 0.05); the expression of TGF-α in the liver tissues was significantly increased at the 4th week (P < 0.05); the expression of TGF-β1 in the liver tissues was gradually significantly increased in every model group (P < 0.05).

CONCLUSIONIn the formation process of cirrhosis, the expression of TGF-α was increased in liver of cirrhosis group at the 4th week, and later it was suppressed; the expression of TGF-β1 was continuously increased. The characteristic dynamic changes of TGF-α and TGF-β1 might be related to sustained endotoxemia, the high level of TNF-α and hyperhomocysteinemia.

Alanine Transaminase ; blood ; Animals ; Carbon Tetrachloride ; Endotoxins ; blood ; Homocysteine ; blood ; Liver Cirrhosis ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor alpha ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; metabolism

OBJECTIVETo establish a new method of multiplex semi-nested polymerase chain reaction (PCR) to detect pathogens in cerebrospinal fluid (CSF).

METHODSAccording to the analysis of the conservative and variable regions in bacterial 16S rRNA genes, we designed universal primers for all bacteria and specific primers for most gram-positive and gram-negative bacteria. All primers were added into the same reaction systems successively of a two-step PCR assay to amplify the different bacterial DNA in CSF, and the results were compared with common culture method with sensitivity and the specificity both detected at the same time.

RESULTSBoth gram-positive and gram-negative bacteria amplified DNA fragment about 1,032 bp after first-step amplification with universal primers. In the second step, specific fragments of 336 bp and 127 bp were amplified in gram-positive and gram-negative bacteria respectively besides fragments of 1,032 bp; The detection limit for E. coli was 8 cfu/ml. The comparison of 62 CSF samples detected by both multiplex semi-PCR and conventional culture method revealed sensitivity, specificity, positive and negative values of 93.8%, 95.7%, 88.2%, and 97.8% respectively for PCR.

CONCLUSIONThe result suggested that the multiplex semi-nested PCR we established was sensitive, specific and rapid method for clinical laboratory to detect pathogens in CSF.

Cerebrospinal Fluid ; microbiology ; DNA Primers ; genetics ; DNA Probes ; genetics ; DNA, Bacterial ; cerebrospinal fluid ; isolation & purification ; Escherichia coli ; genetics ; isolation & purification ; Gram-Negative Bacteria ; genetics ; isolation & purification ; Gram-Positive Bacteria ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; methods ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Sensitivity and Specificity ; Staphylococcus aureus ; genetics ; isolation & purification

Objective To observe the clinical features of atrial fibrillation (AF) patients, and to explore the correlation between the routine detection index and the new-onset AF and to find out risk factors for new AF in critically ill patients. Methods A prospective observational study was conducted. The patients with AF admitted to intensive care unit (ICU) of the Affiliated Hospital of Guizhou Medical University from March 2016 to June 2017 were enrolled. The patients were divided into new-onset AF group and past-existed AF group according to their past history of AF (including persistent AF, paroxysmal AF or permanent AF). In addition, patients in ICU without history of AF and new-onset AF were selected as the control group (no AF group). The general epidemiological characteristics of patients in three groups, and the blood biochemical, coagulation and other related indicators at the time of AF occurred (new-onset AF group) or 48 hours after ICU admission (AF group and no AF group) were analyzed; the difference of laboratory indexes between patients in new-onset AF group with AF within 48 hours before occurred and patients in no AF group within 48 hours after admission to ICU was compared. The relationship between each index and new-onset AF were analyzed. Pearson or Spearman rank correlation was used for analysis. Risk factors of new-onset AF were analyzed by Logistic regression analysis. Results 1 673 patients were admitted to ICU, including 179 cases of AF (10.70%), and 106 males and 73 females, with an average age of (71.73±23.22) years. There was 75 new-onset AF (morbidity 4.48%), and had a 28-day mortality of 45.33% (34/75). There were differences in age, previous heart disease and heart failure (HF) among new-onset AF group (n = 75), past-existed AF group (n = 104) and no AF group (n = 75). Compared with other two groups, renal insufficiency rates, troponin, serum sodium, calcium and procalcitonin levels were higher, mechanical ventilation time and the length of ICU stay were significantly prolonged, ICU and hospitalization costs were higher in new-onset AF group. Compared with no AF group, new-onset AF patients with the higher percentage of septic shock, the accumulation of vascular contraction drugs within 24 hours after AF usage were higher, and used more anti-arrhythmic drugs, has higher brain natriuretic peptide (pro-BNP), serum creatinine, blood lactic acid levels, and lower albumin, oxygenation index, and serum potassium levels, sequential organ failure assessment (SOFA) score, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score and 28-day mortality were higher. Correlation analysis showed that age, APACHE Ⅱ score, septic shock, HF, cardiovascular disease, renal insufficiency were positively correlated with new-onset AF (r values were 0.393, 0.270, 0.386, 0.251, 0.194, 0.170;P values were 0.000, 0.001, 0.000, 0.002, 0.017, 0.037, respectively). The age [odds ratio (OR) = 0.962, P = 0.046], basic oxygenation index (OR = 1.005, P = 0.028) and serum potassium levels (OR = 1.638, P = 0.022) were the risk factors for new-onset AF. Conclusions Critical patients with a high incidence of AF, new-onset AF significantly prolong the length of ICU stay; age, APACHE Ⅱ score, septic shock, cardiovascular disease, and renal insufficiency are related to new-onset AF; age, basic oxygenation index and serum potassium levels are risk factors for new-onset AF.

The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.
Chemistry, Pharmaceutical ; methods ; Ginsenosides ; chemistry ; Mass Spectrometry ; Panax ; chemistry ; growth & development ; Plant Extracts ; chemistry ; Plant Roots ; chemistry

OBJECTIVETo study the protective effects of garlic oil (GO) on the peripheral nerve injuries induced by n-hexane.

METHODSMale Wistar rats were randomly divided into four groups (10 rats in each group): the control, the n-hexane treatment (2000 mg/kg), the low dose GO, and the high dose GO groups. The rats in the low and high doses of GO groups were pretreated with GO (40 and 80 mg/kg) before exposure to n-hexane (2000 mg/ kg), while the animals of the n-hexane treatment group were given normal saline and then 2000 mg/ kg n-hexane. The rats were exposed to GO and n-hexane 6 times a week for 10 weeks. The gait scores and staying time on the rotating rod for all rats were detected every two weeks. The rats were sacrificed at the end of ten weeks, then the levels of alcohol dehydrogenase (ADH), maleic dialdehyde (MDA), reduced glutathione (GSH), glutathione peroxidase(GSH-Px), total antioxidation capacity(T-AOC) and the ability of inhibition of *OH in livers were examined.

RESULTSThe gait scores increased significantly and the time staying on the rotating rod obviously decreased in rats of n-hexane treatment group, as compared with control group (P < 0.05 or P < 0.01). In the hepatic tissues of n-hexane group, the levels of MDA and ADH significantly increased, the activities of GSH-Px, T-AOC and the ability of inhibition of *OH obviously decreased, as compared to control group (P < 0.05 or P < 0.01). In 2 GO groups, the gait scores and the staying time on the rotating rod were significantly improved, the levels of MDA and ADH significantly decreased, the activities of GSH-Px, T-AOC and the ability of inhibition of *OH obviously increased, as compared with n-hexane group (P < 0.05 or P < 0.01 ).

CONCLUSIONADH could play an important role in the protective effects induced by garlic oil on the peripheral nerve injuries produced by n-hexane.

Alcohol Dehydrogenase ; metabolism ; Animals ; Garlic ; Hexanes ; toxicity ; Lipid Peroxidation ; drug effects ; Liver ; enzymology ; metabolism ; Male ; Neuroprotective Agents ; pharmacology ; Peripheral Nerve Injuries ; chemically induced ; metabolism ; Plant Oils ; pharmacology ; Rats ; Rats, Wistar