Objective To investigate the effect of selenium on proliferation and apoptosis of chondrocytes of articular cartilage cultured in vitro in Kaschin-Beck disease(KBD) patients and normal person, to explore the role of selenium in control of KBD, and to provide evidence for selenium's effect on the growth of normal cartilage cells. Methods The articular cartilage samples of grade Ⅱ and Ⅲ KBD patients were selected according to the national "Clinical Diagnosis of KBD" (GB 16003-1995). Chondrocytes of 5 KBD and 5 non-endemic normal accidentswere separated and cultured in vitro. KBD group and control group were given different doses of selenium (0,0.0125,0.0250,0.0500,0.1000,0.2500,0.5000,1.0000 mg/L, respectively). Methyl thiazolyl tetrazolium (MTT),flow cytometric analysis, and immunocytochemical staining were used to observe the effect of selenium on cell growth and apoptosis in KBD and normal persons. Results MTT results showed that the cell proliferation rate in each dosage group of the control group at the 6th day(0.086 ± 0.025,0.077 ± 0.012,0.073 ± 0.027,0.071 ± 0.017,0.058 ± 0.028,0.052 ± 0.028 and 0.046 ± 0.037) was significantly lower than that of 0 mg/L group(0.138 ± 0.026,all P ＜ 0.05);the average cell proliferation rate was negative( - 0.001 ± 0.001, - 0.003 ± 0.000, - 0.003 ± 0.001and - 0.004 ± 0.001 ) in 0.1000 - 1.0000 mg/L dose group, which was significantly lower than that of the 0 mg/L group(0.025 ± 0.003, all P ＜ 0.05);compared with 0 mg/L group(0. 115 ± 0.011), the KBD 0.2500 mg/L dose group promoted cell proliferation(0.128 ± 0.037, P ＜ 0.05), the KBD 1.0000 mg/L dose group inhibited cell growth (0.071 ± 0.019, P ＜ 0.05). The apoptotic rate of 0.0500 - 1.0000 mg/L dose control group [ (18.88 ± 0.02)%,(17.58 ± 0.01)%, (17.09 ± 0.04)%, (56.00 ± 0.02)%, (57.85 ± 0.03)% ] were higher than that of the 0 mg/L group[(13.51 ± 0.01)%, all P ＜ 0.05];compared with 0 mg/L group[(25.84 ± 0.02)%], the apoptotic rate in KBD 0.0250 - 0.2500 mg/L dose group [ ( 13.69 ± 0.02) %, ( 15.96 ± 0.03 ) %, ( 16.68 ± 0.03 ) %, ( 16.67 ± 0.02) % ]were lower, and the apoptotic rate in 0.5000, 1.0000 mg/L dose group [ (59.58 ± 0.03)%, (73.48 ± 0.04)% ] were significantly higher(all P ＜ 0.05). The Fas expression in KBD 0.0500 - 0.2500 mg/L dose groups[ (41.2 ± 1.5)%,(40.3 ± 2.0)%, (50.2 ± 2.5)%] were lower than those of the same dose control group with selenium intervention [(52.4 ± 1.0)%, (67.2 ± 4.0)%, (75.1 ± 5.0)%, all P ＜ 0.05], the caspase-3 expression in KBD 0.0500,0.1000 mg/L dose groups[ (40.8 ± 1.1 )%, (45.1 ± 2.1 )%] were lower than those of the same dose control group with selenium intervention[ (68.0 ± 3.0)%, (70.6 ± 3.5)%, all P ＜ 0.05 ]. Conclusions Appropriate dose of selenium supplementation (0.1000 - 0.2500 mg/L) could promote the growth of KBD chondrocyte, decrease cell apoptosis,but have a damage when the dose of selenium ＞ 0.5000 mg/L;doses of selenium that could promote the growth of KBD chondrocyte does not mean to promote the growth of normal cartilage cells in vivo.

Objective To investigate the effect of growth inhibition and apoptosis induction of sunitinib-liposome-loaded microbubbles on renal carcinoma cell strain.Methods GRC-1 cell strain was cultured in vitro,and was divided into 6 groups:blank control group,pure microbubbles group,pure lipsomes group,sunitinib group,sunitinib-liposome-loaded microbubbles without ultrasound treat group,sunitinib liposome-loaded microbubbles with ultrasound treat group.Growth inhibition in different groups was observed at different time with MTT assay,apoptosis induction with Sigma-FlTC technology and transmission electron microscope.Results The growth inhibition and apoptosis promotion of GRC-1 cell were significantly increased in sunitinib-liposome-loaded microbubbles with ultrasound treat group compared to the other groups.Conclusions Microbubble guided sunitinih delivery can increase the effect of the growth inhibition and apoptosis induction of GRC-1 cells,which may provide an more effective approach for cancer treatment.

This study is to study is to investigate the coumarins from Fruit of Cnidium monnieri and their cytotoxic activities. The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for their cytoxic activities by MTT method. Eleven compounds were isolated and identified as osthole (1), bergaptan (2), xanthotoxol (3), xanthotoxin (4), imperatorin (5), isopimpinellin (6), osthenol (7), psoralen (8), 5,7-dimethoxycoumarin (9), oxypeucedaninhydrate (10), and swietenocoumarin F (11). Compounds 7, 9-11 were isolated from the Cnidium genus for the first time. Compounds 1,5,10 and 11 showed significant cytotoxic activities against L1210 cell lines at a concentration of 1 x 10(-5) mol x L(-1) with inhibitory rates of were 70.13, 63.10, 55.77, and 75.08% respectively.
Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cnidium ; chemistry ; toxicity ; Coumarins ; chemistry ; isolation & purification ; toxicity ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Fruit ; chemistry ; toxicity ; Mice ; Molecular Structure

Objective To observe the clinical effect and safety of Wenxin formula in the treatment of stable coronary heart disease angina patients with Syndrome of Yang deficiency with stagnation of the Blood and retention of Phlegm (SYBP). Methods A total of 65 stable coronary heart disease angina patients with SYBP were randomly assigned to the trial group and the control group (34 in the trial group and 31 in the control group). The trial group was treated with Wenxin formula, while the control group was treated with atorvastatin. The treatment lasted 4 weeks. The therapeutic effects on angina, reduction of Nitroglycerin, changes of electrocardiogram (ECG) and TCM syndrome score were observed before and after treatment.ResultsAfter treatment, the trial group had a similar effect with the control group in the aspects of the therapeutic effects on angina, reduction of Nitroglycerin and the ECG change. There was no statistical difference between the two groups (P>0.05). The changes of TCM syndrome score in the trial group was superior to the control group (P<0.05). The total effective rates of the trial group and the control group were 88.2% and 74.2%. The significant efficiency of the treatment group (29.41%) was significantly superior to the control group (9.68%) (P<0.05). No adverse reaction occurred during the therapeutic course.Conclusions The Wenxin formula was safe and effective in the treatment of stable coronary heart disease angina patients with SYBP.

OBJECTIVETo observe the clinical efficacy on tic disorders (TD) treated with local acupuncture.

METHODSThe entire data of 196 cases, of which, 114 cases in acupuncture group were treated mainly with local acupuncture and 82 cases in western medication group were taken Tiapride tablets orally. YGTSS method was adopted for scoring and assessing the clinical efficacy.

RESULTSThe apparent improvement was achieved in YGTSS motor tic score in two groups. The markedly effective rates were 90.4% and 84.2% in acupuncture group and western medication group, respectively (P < 0.05), without significant difference in statistics. The effective rates of transient tic disorder (TTD) in acupuncture group and western medication group were 100.0% and 83.3%, respectively (P < 0.05), which indicated that the efficacy of acupuncture on TTD was advantageous to medical therapy. The effective rates of three kinds of TD in acupuncture group were 100.0%, 88.2% and 84.2% separately, of which, the efficacy of acupuncture on TTD was advantageous as compared with that on chronic tic disorder (CTD) and Toureett's syndrome (TS) (P < 0.05).

CONCLUSIONThe overall efficacy on TD is similar between acupuncture and medical therapy, but the clinical efficacy of acupuncture on TTD is superior to medical therapy. Meanwhile, the efficacy of acupuncture on TTD is advantageous to that on CTD and TS. Hence, the intervention at early time can achieve satisfactory efficacy and probably prevent from CTD or TS.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Tic Disorders ; therapy

OBJECTIVETo prepare highly specific anti-ouabain polyclonal antibody for detecting endogenous ouabain in tissues.

METHODSOuabain-BSA compound was used to immunize hens, and the eggs were collected one week after the first immunization. The IgY antibodies in the egg yolk were separated and purified by PEG-6000 Method, and analyzed by 12% SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) for titration. The IgY antibodies obtained were applied subsequently in ELISA and immunohistochemistry.

RESULTSThe IgY titer increased rapidly after the second immunization, with the highest titer of 1:10240 that lasted for at least 4 weeks. Competitive ELISA for IgY detection showed an average intraassay coefficient of variation (CV) of 2.03% and an inter-assay CV of 2.34%. Immunohistochemistry visualized the location of the endogenous ouabain mainly in the cytoplasm of the zona reticularis of rat adrenal cortex.

CONCLUSIONImmunization of hens allows efficient preparation of IgY antibody which can be used in routine immunoassays.

Animals ; Calibration ; Cattle ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Immunization ; methods ; Immunoglobulins ; immunology ; Immunohistochemistry ; Ouabain ; analysis ; immunology ; Rats

To analyze and evaluate predictive value of preoperative changes of serum levels of brain natri‐uretic peptide (BNP) and homocysteine (Hcy) levels for therapeutic effect and prognosis of coronary artery bypass grafting (CABG ) in patients undergoing CABG .Methods : Clinical data of 115 patients undergoing CABG in our hospital were collected .Preoperative serum levels of BNP and Hcy were compared among patients with different disease severity , therapeutic effect and prognosis .Influence of serum BNP and Hcy levels on therapeutic effect and prognosis of CABG were analyzed .Results : Compared with medium and mild disease group , there were significant rise in preoperative serum levels of BNP [ (151.86 ± 22.57) pg／L vs.(82.57 ± 10.26) pg／L vs.(283.51 ± 32.47) pg／L] and Hcy [ (18.37 ± 4.51) μmol／L vs.(12.74 ± 2. 04) μmol／L vs.(31.56 ± 5.17) μmol／L] in severe disease group , and those of medium disease group were significantly higher than those of mild disease group , P＝0.001 all ;compared with effective group and ineffective group , there were significant reductions in preoperative serum levels of BNP [ (227. 49 ± 24. 52) pg／L vs.(308.26 ± 34.12) pg／L vs.(90.13 ± 10. 75) pg／L] and Hcy [ (29. 12 ± 5. 83) μmol／L vs.(46.15 ± 7.49) μmol／L vs.(19.03 ± 3.77) μmol／L] in markedly effective group , and those of effec‐tive group were significantly lower than those of ineffective group , P＝ 0.001 all.Compared with stenotic graft group and smooth graft group , there were significant rise in preoperative serum levels of BNP [ (271. 47 ± 25.18) pg／L vs .(92.41 ± 11.06) pg／L vs.(312. 54 ± 35.06) pg／L] and Hcy [ (33.08 ± 6. 14) μmol／L vs.(20. 05 ± 3. 68) μmol／L vs.(50.21 ± 7.75 ) μmol／L ] in death group , and those of stenotic graft group were significantly higher than those of smooth graft group , P＝0.001 all .Spearman correlation analysis indicated that serum BNP and Hcy levels were significant positively correlated with severity of CHD and prognosis of CABG ( r＝0. 624～0.814 , P＜0.05 or ＜0. 01).Multifactor Logistic regression analysis indicated that serum BNP and Hcy levels were independent risk factors for prognosis after CABG in CHD patients (OR＝ 5.133 , 1. 803 , P＝ 0.001 both ).Conclusion : The higher preoperative levels of BNP and Hcy before CABG are , the worse therapeutic effects are , and the higher risk of adverse prognosis of vascular stenosis and death are .

BACKGROUNDThe 2009 influenza A (H1N1) virus infection is associated with the high risk of severe complications and is spreading more rapidly throughout the world than other reported seasonal influenzas. This study aimed to evaluate the efficacy and safety of the nature herbal medicine Lianhuaqingwen capsule (LHC) in patients infected with influenza A (H1N1) virus.

METHODSA total of 244 patients aged 16 - 65 years confirmed with influenza A (H1N1) virus infection by the real time RT-PCR were randomized to one of two treatment groups of 122 patients each. Each group assigned to receive either LHC or Oseltamivir for five days and observation for seven days. The patients were enrolled within 36 hours of illness onset if they had an axillary temperature of ≥ 37.4°C and with at least one of the following symptoms: nasal obstruction, runny nose, cough, sore throat, fatigue, headache, myalgia, chills and sweating. The primary end point was the duration of illness.

RESULTSOf 244 patients, 240 (98.36%) patients with a median age 21 years completed the study between October 24, 2009 and November 23, 2009. There were no significant overall differences between LHC treated and Oseltamivir treated patients in the median duration of illness (LHC 69 hours vs. Oseltamivir 85 hours P > 0.05) or the median duration of viral shedding (LHC 103 hours vs. Oseltamivir 96 hours, P > 0.05). However, it was worthwhile to note that LHC significantly reduced the severity of illness and the duration of symptoms including fever, cough, sore throat, and fatigue (P < 0.05). Both study medications were well tolerated. No drug related serious adverse events occurred during the study.

CONCLUSIONSCompared with Oseltamivir, LHC achieved a similar therapeutic effectiveness reduction of the duration of illness and duration of viral shedding. Therefore, LHC might be an alternative therapeutic measure for influenza A (H1N1) virus infections.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; pathogenicity ; Influenza, Human ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVERNA interference is a new technology that inhibit effectively the expression the specific genes. The current study was designed to investigate whether the plasmid containing the short hairpin RNA (shRNA) of angiotensin II type 1 receptor (AT(1)R) can inhibit the hyperplasia of vascular smooth muscle cells in rat.

METHODSThe plasmids containing the shRNA of AT(1)R were constructed, and transfected vascular smooth muscle cell (VSMC) to detect the effect on the AT(1)R expression by RT-PCR and Western blot, observe the shape of VSMCs by the inverted phase contrast microscope, and detect the hyperplasia of VSMCs by trypan blues staining and MTT.

RESULTSThe plasmids was certified to be in the right rank. After transfecting cells, there was significant difference (P < 0.01) in the expression of AT(1)R mRNA between the plasmid transfected group (pAT(1)R-shRNA(1) 1.37 +/- 0.15; pAT(1)R-shRNA(2) 1.45 +/- 0.12) and the control group (2.09 +/- 0.26), and there was significant difference (P < 0.01) in the expression of AT(1)R protein between the gene transfected group (pAT(1)R-shRNA1 1.12 +/- 0.04; pAT(1)R-shRNA2 1.20 +/- 0.07) and the control group (3.17 +/- 0.21). It is shown that pAT(1)R-shRNA can decrease the expression of AT(1)R mRNA and protein. There was significant difference (P < 0.01) in the Cell number between the plasmid transfected adding AngII group (pAT(1)R-shRNA1 5.48 +/- 0.44; pAT(1)R-shRNA2 5.55 +/- 0.45) and the AngII control group (8.13 +/- 0.41); there was significant difference (P < 0.01) in the Ratio of light density by MTT between the plasmid transfected adding AngII group (pAT(1)R-shRNA1 0.365 +/- 0.024; pAT(1)R-shRNA2 0.307 +/- 0.025) and the control group (0.485 +/- 0.011); It is shown that that pAT(1)R-shRNA can inhibit the hyperplasia of VSMCs, and matching the result of morphology observation.

CONCLUSIONSThe plasmids containing the shRNA of AT(1)R can inhibit the expression of AT(1)R mRNA and protein in VSMCs, and inhibit the hyperplasia of VSMCs induced by AngII in rat.

Angiotensin II ; pharmacology ; Animals ; Aorta ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hyperplasia ; Muscle, Smooth, Vascular ; pathology ; Myocytes, Smooth Muscle ; cytology ; metabolism ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; biosynthesis ; genetics ; Transfection

OBJECTIVETo observe the injury on micro-skin induced by a self designed micro-skin machine.

METHODSMicro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.

RESULTSTransmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).

CONCLUSIONThe cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.

Burns ; surgery ; Cell Division ; Epithelium ; pathology ; Humans ; Microscopy, Electron ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Wound Healing