Cooperative Behavior ; Depression ; Humans ; International Cooperation ; Research

OBJECTIVETo explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment.

METHODThe three kinds of explant of bud, leaf and calli were induced by colchicine treatment.

RESULTThe induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different.

CONCLUSIONInduction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

Chromosomes, Plant ; genetics ; Colchicine ; pharmacology ; Plant Leaves ; anatomy & histology ; genetics ; growth & development ; Plant Shoots ; anatomy & histology ; genetics ; growth & development ; Plants, Medicinal ; anatomy & histology ; genetics ; growth & development ; Polyploidy ; Salvia ; anatomy & histology ; genetics ; growth & development

OBJECTIVETo study the leptin resistance mechanism of Xiaoyan Decoction (XD) in lung cancer cachexia (LCC) rats.

METHODSAn LCC rat model was established. Totally 40 rats were randomly divided into the normal control group, the LCC model group, the XD group, and the positive control group, 10 in each group. After LCC model was set up, rats in the LCC model group were administered with normal saline, 2 mL each time. Rats in the XD group were administered with XD at the daily dose of 2 mL. Those in the positive control group were administered with Medroxyprogesterone Acetate suspension (20 mg/kg) by gastrogavage at the daily dose of 2 mL. All medication lasted for 14 days. The general condition and tumor growth were observed. Serum levels of leptin and leptin receptor in the hypothalamus were detected using enzyme-linked immunosorbent assay. Contents of neuropeptide Y (NPY) and anorexia for genomic POMC were detected using real-time PCR technique.

RESULTSSerum leptin levels were lower in the LCC model group than in the normal control group with statistical significance (P < 0.05). Compared with the LCC model groups, serum leptin levels significantly increased in the XD group (P < 0.01). Leptin receptor levels in the hypothalamus increased significantly in the LCC model group (P < 0.01). Increased receptor levels in the LCC model group indicated that either XD or Medroxyprogesterone Acetate could effectively reduce levels of leptin receptor with statistical significance (P < 0.01). There was also statistical difference between the XD group and the positive control group (P < 0.05). Contents of NPY was higher in the LCC model group than in the other groups with statistical difference (P < 0.05). There was no statistical difference in NPY between the normal control group and the rest 2 treatment groups (P > 0.05). There was statistical difference in POMC between the normal control group and the LCC model group (P < 0.05). POMC could be decreased in the XD group and the positive control group with statistical significance (P < 0.05), and it was more obviously decreased in the XD group (P < 0.05).

CONCLUSIONSLeptin resistance existed in LCC rats. XD could increase serum leptin levels and reduce leptin receptor levels in the hypothalamus. LCC could be improved by elevating NPY contents in the hypothalamus and reducing POMC contents, promoting the appetite, and increasing food intake from the periphery pathway and the central pathway.

Animals ; Cachexia ; drug therapy ; etiology ; Drugs, Chinese Herbal ; therapeutic use ; Eating ; Humans ; Hypothalamus ; metabolism ; Leptin ; metabolism ; Lung Neoplasms ; complications ; Neuropeptide Y ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

DNA, Ribosomal ; genetics ; Genome, Plant ; Phylogeny ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; genetics ; RNA, Ribosomal, 5S ; genetics ; Sequence Homology, Nucleic Acid ; Species Specificity ; Valerian ; classification ; genetics

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.

Objective To explore the significance of 18 F-fluorodeoxyglucose positronemission tomography/ computed tomography (18F-FDG PET/CT) scanning for childhood neuroblastoma clinical staging.Methods From December 2014 to December 2015,59 patients confirmed as NB in Beijing Children's Hospital,Capital Medical University,and finding or clinical features were selected by histopathology.Those patients underwent 18 F-FDG PET/CT,bone scan,cranial magnetic resonance image (MRI),bone marrow puncture (two sites) and biopsy,regional CT,ultrasound,serum tumor markers like lactic dehydrogenase and neuronspecific enolase.In comparison of 18F-FDG PET/CT image findings with other detective methods,the differences were analyzed between the primary sites and metastasis were analyzed.Results (1) General features:Out of 59 children with NB,31 were males,28 were females.Primary lesions were as follows:14 cases located in the postmediastinum,44 cases in the retroperitoneum (1 case in retroperitoneum + postmediastinum),1 case in left neck.International Neuroblastoma Staging System Ⅰ to Ⅳ:O,1,9 and 49 cases,respectively.(2)18F-FDG PET-CT manifestations:The maximum of standardized uptake value (SUVmax):before-treatment group was (2.34 ± 1.06) which was larger than the post-surgery group value of (1.08 ± 0.50),and the difference was significant(F =5.699,P =0.000);bone marrow metastasis ranged from 1.5 to 2.9,regional lymphatic metastasis ranged from 1.0 to 2.1.(3)18F-FDG PET-CT imaging compared with other detection:in bone metastases,the whole body bone scan finding were normal in 3 cases,while PET/CT showed disseminated bone and bone marrow involvement and bone scanning showed 2 cases had metastases,combined with 18 F-FDG PET/CT,which were considered as postoperative inflammatory reaction or residual tumor tissues invade adjacent intervertebral foramen.Bone cytology was positive in 30 cases,while PET/CT showed 34 patients with bone marrow metastasis,and 1 case was suspicious.In central metastasis,1 case of PET/CT showed epidural metastasis,while cranial MRI was negative;Cranial MRI showed 9 cases had skull metastasis,1 case had orbital metastasis,1 case had meningeal metastases;while 18F-FDG PET/CT showed no abnormality.All children had no parenchymal metastasis.Corresponding tumor markers as LDH was related to the maximum value of primary tumor focal SUVmax (rs =0.581,P ＜ 0.01).Conclusions 18 F-FDG PET/CT can fully display the distribution of primary sites and metastases,and can be served as medical imaging evidence for both the diagnosis and staging of neuroblastoma.But as for cranial bone and central metastasis,it has high false negative rate.Clinical practice should be combined with cranial MRI to improve relevance ratio.

AIM:To investigate the effect of sodium hydrosulfide(NaHS)on cardiac function and activity of renin-angiotensin(Ang)-aldosterone(ALD)system(RAAS)in the rats with chronic heart failure(CHF).METHODS:The CHF rat model was established by abdominal aortic coarctation.SD rats were randomly divided into sham operation group,model group,low dose of NaHS group and high dose of NaHS group(n=6).The left ventricular end-diastolic di-ameter(LVEDD), left ventricular end-systolic diameter(LVESD)and left ventricular ejection fraction(LVEF)were measured before and after treatment by echocardiography in each group.The levels of renin,AngⅡand ALD in the plasma were measured by ELISA.The expression of angiotensin-converting enzyme(ACE)and angiotensin Ⅱ type 1 receptor (AT1R)at mRNA and protein levels in the myocardium tissues was determined by qPCR and Western blot,respectively. RESULTS:After treatment with NaHS,compared with model group and before treatment,LVEDD and LVESD in low dose of NaHS group and high dose of NaHS group were decreased significantly, while LVEF was increased significantly(P<0.05).Compared with low dose of NaHS group,LVEDD and LVESD were decreased,while LVEF was increased in high dose of NaHS group(P<0.05).Compared with sham operation group,the levels of renin,AngⅡand ALD in the plasma of model group were significantly increased(P<0.05),and the expression of ACE and AT1R at mRNA and protein levels in the myocardium tissues of model group were significantly increased(P<0.05).Compared with model group,the plas-ma levels of renin,AngⅡand ALD in low dose of NaHS group and high dose of NaHS group were significantly decreased (P<0.05),and the myocardial expression of ACE and AT 1R at mRNA and protein levels was significantly down-regulated (P<0.05).The plasma levels of renin,AngⅡand ALD,and the myocardial expression of ACE and AT 1R at mRNA and protein levels in high dose of NaHS group were significantly lower than those in low dose of NaHS group(P<0.05). CONCLUSION:NaHS inhibits the activation of RAAS,thus improving the cardiac function of CHF rats,and the effect of high-dose NaHS is better than that of low-dose NaHS.

The "four-in-one" approach is based on the four-dimensional perspective of "property, position, tendency and syndrome", which helps to identify and analyze classical prescriptions in a multi-dimensional and three-dimensional way. The early pathogenesis of chronic heart failure (CHF) is deficiency of heart Qi and heart Yang and disorder in Qi transformation in triple energizer, while in the later stage of the disease, it progresses from deficiency to excess, with simultaneous occurrence of deficiency and excess syndromes. Fuling Guizhi Baizhu Gancao Decoction (Linggui Zhugan Decoction) plays its role in treating chronic heart failure by the four elements of "property, position, tendency and syndrome". Property—Linggui Zhugan Decoction is pungent, sweet, slight sweet and bitter in flavor, but warm in property. The sweet is able to tonify deficiency; the pungent is responsible for dispersing Yang, promoting Qi and draining water retention; the warm nourishes the spleen, raises Yang Qi and resolves phlegm; the bitter could excrete diuresis and dry dampness to guarantee the smooth operation of three energizer. Position-Linggui Zhugan Decoction acts on the heart, spleen and triple energizer. It can stimulate heart Yang, strengthen the spleen, resolve phlegm, and regulate the waterways to promote the Qi transformation in triple energizer. Tendency-The tendency of Linggui Zhugan Decoction is upward and downward in parallel, both internal and external. Warming up and promoting diuresis, raising Yang up and tonifying deficiency, it is conducive to the Yang Qi transformation in triple energizer. Syndrome-Linggui Zhugan Decoction is indicated for the syndrome of heart Yang deficiency and water-fluid retention, which begins with the upper abdomen swelling, Qi rushes against the chest. It is widely used in the treatment of water-vapor impulse heart disease. The disorder of Qi transformation in triple energizer is the main mechanism of recurrent CHF. Linggui Zhugan Decoction can not only warm the fire of Qi transformation in triple energizer, but also smooth the pathway of Qi transformation in triple energizer, which is compatible with the treatment of systemic fluid retention in chronic heart failure. Its pharmacological mechanisms include anti-inflammation, anti-platelet aggregation, regulation of cardiomyocyte cell membrane ion channels, protection against ischemia-reperfusion injury and modulation of vasodilation, etc. Deconstructing Linggui Zhugan Decoction with "four-in-one" approach and discussing its mechanism for treating CHF in combination with the theory of "Qi transformation in triple energizer", have great significance to rejuvenate the vitality of classical prescriptions and to apply them accurately and effectively.

OBJECTIVETo investigate the protective effect of hepatocyte growth factor (HGF) on protein synthesis in rat cardiomyocytes exposed to gamma-ray irradiation.

METHODSPrimary cultured cardiomyocytes were irradiated with single-dose (20 Gy) gamma ray in the absence or presence of HGF (40 ng/ml) added in the cell culture 3 h before the exposure. Forty-eight hours after irradiation, the total cellular protein was measured and cell cycle analyzed by flow cytometry. The cardiomyoctes were also infected with AdGFP 48 h after irradiation and the fluorescence intensity of the green fluorescence protein (GFP) in the cells determined by flow cytometry 48 h after infection.

RESULTSThe protein synthesis was decreased significantly in the irradiated cardiomyocytes as compared with the control group (P<0.01), but was remedied significantly by incubation of the cells with HGF before the exposure (P<0.05). Flow cytometry revealed much lower mean fluorescence intensity (MFI) of GFP in irradiated cardiomycytes than in cells without the exposure (P<0.01); The MFI was higher in HGF-treated cardiomyocytes than in cells without HGF treatment following the exposure (P<0.01).

CONCLUSIONGamma ray irradiation inhibits protein synthesis in cardiomyocytes, and HGF may attenuate this effect of gamma ray exposure for cardiomyocyte protection.

Animals ; Animals, Newborn ; Cell Cycle ; drug effects ; radiation effects ; Cells, Cultured ; Flow Cytometry ; Gamma Rays ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatocyte Growth Factor ; pharmacology ; Microscopy, Fluorescence ; Myocytes, Cardiac ; cytology ; metabolism ; Protein Biosynthesis ; drug effects ; radiation effects ; Rats ; Rats, Wistar