Objective To study the inducement of U251 glioblastoma cell apoptosis in vivo through up-regulating PUMA expresion and knocking down miR-221/222, and explore its mechanism.Methods Nude mouse xenograft models were established in 5-week-old BALB/c nude mice by subcutaneous vaccination of U251 glioblastomas; 1 week later, they were treated with intratumoral injection of lipofcctamine-mediated miRNA-221/222 antisense oligonucleotides (GroupA), nonsense sequences (Group B) and controls (Group C),respectively (n=8).The tumor growth was monitored until the end of observation period (28 d after the treatment) and pathological changes of the glioblastoma tissues were observed by HE staining at the end of observation.Fluorescence in situ hybridization (FISH) and real-time PCR were employed to measure the miR-221 and miR-222 expressions. Terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL) assay was used to detect the apoptosis of glioblastomas.Immunohistochemistry and Westem blotting were used to detect the expressions of PUMA,bax,bcl-2 and p53 in removed tumor specimens. Results The volume in Group A was significantly smaller than that of those in group B and group C 6-28 dater treatment (P=0.006). The miR-221 and miR-222 mRNA expressions in Group A were significantly decreased as compared with those of those in group B and group C.HE staining indicated that decreased heteromorphism and reduced number of new vessels in Group A were noted as compared with those in group B and group C.The cell apoptotic index in Group A was significantly higher than that in group B and group C (P＜0.05).Immunohistochemistry showed that the expression levels of PUMA and bax in Group A was significantly up-regulated as compared with those in group B and group C, while the expression of bcl-2 in Group A was significantly down-regulated as compared with that in group B and group C; and no significant changes were noted in the p53 expression. Conclusion By up-regulating PUMA expresion,knocking down miR-221/222 can induce U251 glioma apoptosis in vivo.

Phytochemical investigation on the EtOH extract from the aerial part of Callicarpa kwangtungensis led to the isolation and characterization of 10 caffeoyl phenylethanoid glycosides, 2'-acetylacteoside (1), tubuloside E (2), acteoside (3), tubuloside B (4), isoacteoside (5), alyssonoside (6), 2'-acetylforsythoside B (7), brandioside (8), forsythoside B (9), and poliumoside (10). Compound 4 was isolated from the plants of Verbenaceae,and 6 was obtained from the Callicarpa genus, for the first time, while compounds 1, 2, 5 and 7 were firstly reported from the plant.
Caffeic Acids ; chemistry ; isolation & purification ; Catechols ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Ethanol ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Phenols ; chemistry ; isolation & purification ; Verbenaceae ; chemistry

OBJECTIVETo study the constituents with the pain-relieving activity from the stem rind of Daphne giraldii.

METHODThe partition of the ethanol extract and chromatographic separation of the fractions were carried out by the monitoring of anelgesic pharmacological activity. The structures of the isolated compounds were determined by MS and NMR.

RESULTFour compounds were isolated from the pain-relieving fraction. Three of them were identified as diterpenes, gniditrin (1), gnidicin (2) and daphnetoxin (3). Compound 4 was determined as Z-octadecyl caffeate.

CONCLUSIONCompounds 1, 2 and 4 were isolated from the plant for the first time.

Analgesics ; chemistry ; isolation & purification ; pharmacology ; Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Daphne ; chemistry ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

Objective To study the effects of pulsed electromagnetic fields(PEMFs)of 15Hz and lmT on osteoblast differentiation of bone marrow mesenchymal stem cells.Methods The bone marrow mesenchymal stem cells were isolated and cultured in vitro,the third passage cells were harvested and half of them was exposed to PEMFs,another half exposed to sham PEMFs served as the control.The activity of ALP was measured.And the ex- pression of osteoblast and adipogenesis was measured with the semi-quantitative RT-PCR,adipogenic differentiation was observed with the Oil Red O Staining.Results When compared with the control,PEMFs of 15 Hz and 1 mT significantly increased the activity of ALP(P

Objective:To observe the clinical efficacy of auricular point sticking at different points to relieve the pain in arteriovenous fistula puncture. Methods: A total of 42 patients with arteriovenous fistula were randomized into a Shenmen (TF4) group and an Elbow (SF3) group by the random number table method, with 21 cases in each group. After enrolled into different groups, before the dialysis, patients were given auricular point sticking with Wang Bu Liu Xing ( Semen Vaccariae) seeds at Shenmen (TF4) and Elbow (SF3), respectively. Patients were asked to press the seeds themselves for 2 min each time, four times a day, and an additional 5-15 min before the arteriovenous fistula puncture. Intensive pressing was offered during the puncture, 15-20 presses for each time, and the plasters were changed every 2-3 d. The numerical rating scale (NRS) was used to score the pain level one week before and after auricular point sticking. The NRS score was then compared and analyzed. Results: The intra-group comparison showed that the changes of NRS score in both groups were statistically significant after auricular point sticking (both P＜0.05). After the treatment, there was no significant difference in NRS score between the two groups (P＞0.05). Conclusion: Auricular point sticking at Shenmen (TF4) or Elbow (SF3) can effectively relieve the pain of arteriovenous fistula puncture, and these two points have equivalent analgesic effect.

Objective To explore the effect of atorvastatin on cardiac hypertrophy and to determine the potential mechanism involved. Methods Anin vitro cardiomyocyte hypertrophy from neonatal rats was induced with angiotensinⅡ (AngⅡ) stimulation. Before AngⅡ stimulation, the cultured rat cardiac myocytes were pretreated with atorvastatin at different concentrations (0.1, 1, and 10μmol/L). The following parameters were evaluated: the myocyte surface area,3H-leucine incorporation into myocytes, mRNA expressions of atrial natriuretic peptide, brain natriuretic peptide, matrix metalloproteinase 9, matrix metalloproteinase 2, and interleukin-1β, mRNA and protein expressions of theδ/β peroxisome proliferator-activated receptor (PPAR) subtypes. Results It was shown that atorvastatin could ameliorate AngⅡ-induced neonatal cardiomyocyte hypertrophy in the area of cardiomyocytes,3H-leucine incorporation, and the expression of atrial natriuretic peptide and brain natriuretic peptide markedly. Meanwhile, atorvastatin also inhibited the augmented mRNA level of several cytokines in hypertrophic myocytes. Furthermore, the down-regulated expression of PPAR-δ/β at both the mRNA and protein levels in hypertrophic myocytes could be significantly reversed by atorvastatin treatment. Conclusions Atorvastatin could improve AngⅡ-induced cardiac hypertrophy and inhibit the expression of cytokines. Such effect might be partly achieved through activation of the PPAR-δ/β pathway.

Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus(HBV). Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northem blot, and strong signals could be seen from day 7 to day 14 post-in-fection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably repli-cate and express in HBV-infected tupaia hepatocytes.

OBJECTIVETo study the inhibitory effect and reasons of liposomes survivin antisense oligonucleotides (ASODN) on growth of human gastric carcinoma transplanted subcutaneously in nude mice.

METHODSHuman gastric carcinoma transplanted subcutaneously in nude mice model was established, and subsequently was divided randomly into six groups: control group, liposome group, sense oligonucleotide (SODN) group, 100, 200 and 400 nmol/L ASODN group. Different treatments were given respectively. The weight and volume of subcutaneous tumors were measured, and tumor growth inhibitory rate and decreased rate was calculated. The morphological changes of transplanted tumor cells were observed under light microscope. The expression of survivin was detected by immunohistology (SP). Changes of survivin gene transcription and protein expression were determined by western blot and RT-PCR.

RESULTSGrowth of the tumors was significantly inhibited in all ASODN groups as compared with that in the control, liposome and SODN group. The highest growth inhibitory rate in the 400 nmol/L group is 93%. The number of apoptotic cells of ASODN group increased and expression of survivin became weaken under the microscope. Liquified necrosis regions could be seen in 6 cases (6/12) of tumor tissues. The content of survivin mRNA and protein decreased in all survivin ASODN groups. The survivin protein expression of 400 nmol/L group was about 36.8% of the control group.

CONCLUSIONSSurvivin gene ASODN can inhibit the growth of human gastric carcinoma in nude mice by inducing cells apoptosis and decreasing the expression of survivin mRNA and protein.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Liposomes ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; therapy

OBJECTIVETo investigate the clinical effects of the Chinese traditional medicine Yougui Capsules and Wuziyanzong Pills on sperm viability and motility in patients with oligoasthenospermia.

METHODSA total of 80 infertile men oligoasthenospermia were equally randomized into a trial and a control group, the former treated with Yougui Capsules at 1.68 g tid, while the latter with Wuziyanzong Pills at 6 g bid, both for a course of 12 weeks. The sperm viability and motility of the patients were detected and compared before and after medication.

RESULTSAfter 12 weeks of medication, the sperm viability and percentages of grade a and grade a + b sperm were (65.7 +/- 13.1), (22.5 +/- 9.1) and (47.6 +/- 15.8)% in the trial group, significantly higher than (38.1 +/- 11.1), (13.2 +/- 6.8) and (24.1 +/- 10.9)% in the control (P<0.05). What's more, the above three parameters of the two groups were also significantly higher than those before medication, which were (31.9 +/- 16.9), (8.2% +/- 3.7) and (15.7 +/- 13.9)% in the former and (31.7 +/- 17.0), (7.9 +/- 4.5) and (16.9 +/- 13.6)% in the latter (P<0.05).

CONCLUSIONBoth Yougui Capsules and Wuziyanzong Pills can improve sperm viability and motility in patients with oligoasthenospermia, and the former is even more efficacious than the latter.

Adult ; Asthenozoospermia ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Young Adult

Metabonomics is a new method to study on the metabolic network and the relationship between body and environment, which conforms to the way of traditional Chinese medicine (TCM) research. In the study process of modernization of traditional Chinese medicine, effectively conjunction with metabonomics method will facilitate the integration of TCM with modern biological science and technology, and promote the modernization of TCM. This paper introduce the application of metabonomics in the research of toxicity mechanism of TCM, compatibility mechanism of TCM formula, pharmacology effect of TCM and processing mechanism of TCM. This paper summarize the problems in the TCM metabonomics research and prospect its bright future.
Animals ; Drug Therapy ; Drugs, Chinese Herbal ; adverse effects ; analysis ; metabolism ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; trends ; Metabolomics ; methods ; trends