Background Oxidative stress-induced apoptosis of human lens epithelial cells (LECs) is associated with c-Jun N terminal kinase (JNK) pathway.Quercetin possesses the antioxidation by inhibiting the JNK pathway.However,whether quercetin can protect LECs from the oxygen-induced damage is still not proved.Objective This study attempted to invatigate the effects and its mechanism of quercetin against hyperbaric oxygeninduced LECs apoptosis. Methods Human LECs line SRA01/04 was cultivated and passaged in MEM medium containing 10％ fetal bovine serum and 0.5％ non-essential amino acids for 2 hours,with or without 20 μmol/LSP600125 or 1 μmol/L quercetin prior to exposure to hyperbaric oxygen.Each exposure session remained 6 hours in 99％ O2 and 1％CO2 with a pressure chamber at 588 kPa.The viability of human LECs was detected by MTT.Cell apoptosis was assessed by flow cytometer using Annexin V-FITC apoptosis detection.The expression of JNK/p-JNK,c-Jun/p-c-Jun,caspase 3 and caspase 9 were detected by Western blot. Results LECs viability (A570 ) was 0.835 ±0.082,0.450±0.083,0.654±0.079,0.649±0.090 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.The A570 in the hyperbaric oxygen exposed group was significantly lower than the blank control group ( P =0.000),but those in hyperbaric oxygen + SP600125 group and hyperbaric oxygen+quercetin group were significantly higher than the hyperbaric oxygen exposed group ( P =0.003,0.002 ).The numbers of apoptosis cells were 3.17 ±0.74,19.77 ± 1.44,8.45 ±0.93,7.79 ±0.78 respectively in the blank control group,hyperbaric oxygen exposed group,hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group.Apoptotic LECs were significantly increased in the hyperbaric oxygen exposed group compared with the blank control group ( P=0.000),but those in the hyperbaric oxygen+SP600125 group and hyperbaric oxygen+quercetin group were significantly reduced in comparison with hyperbaric oxygen exposed group (both P=0.000).In additional,expressions of p-JNK,p-c-Jun,caspase 3 and caspase 9 proteins in the cells were elevated in the hyperbaric oxygen exposed group compared with the blank control group (all P =0.000 ),however,those in the hyperbaric oxygen + SP600125 group and hyperbaric oxygen + quercetin group were declined when compared with the hyperbaric oxygen exposed group( all P＜0.05 ). Conclusions JNK pathway is involved in the apoptotic procedure of human LECs induced by oxygen stress.SP600125 and certain concentration of quercetin can interdict the JNK signal pathway and endogenous apoptosis of LECs and further alleviate hyperbaric oxygen-induced damage of LECs.

OBJECTIVETo explore the difference in the efficacy and effect mechanism of subcortical ischemic vascular disease (SIVD) complicated with depression between acupuncture and medication.

METHODSSixty patients were randomized-into an acupuncture group and a medication group, 30 cases in each one. In the acupuncture group, acupuncture was applied to Baihui (GV 20), Shuigou (GV 26), Fengfu (GV 16), Fengchi (GB 20) and the others, once a day, 6 times a week. The treatment of 4 weeks made one session and totally 2 sessions were required. In the medication group, nimodipine 30 mg, three times a day and fluoxetine 20 mg, once a day were prescribed for oral administration, for 8 weeks totally. Before treatment, at the end of the 4th week and at the end of the 8th week of treatment, cerebral blood flow velocity (CBFV) and solubility CD40 ligand (sCD40L) were determined respectively. The scores in Montreal cognitive assessment (MoCA) and Hamilton' s depression scale (HAMD) were evaluated in the two groups. The efficacies on cognitive function and depression symptoms were compared in the patients between the two groups. Results Compared with the outcome before treatment, mean blood flow velocity (Vm) of middle cerebral artery (MCA), anterior cerebral artery (ACA) and posterior cerebral artery (PCA) was increased significantly at the end of the 4th week of treatment in the two groups (all P<0.05). At the end of the 8th week, Vm was increased much significantly (all P<0.01). The differences were not significant in comparison between the two groups (all P>0.05). Compared with the expression before treatment, sCD40L was reduced significantly after treatment in the patients of the two groups (all P<0.01), but the differ- ence was not significant between the two groups (all P>0.05). Compared with that before treatment, MoCA score was increased significantly after treatment in the two groups (all P<0.01), HAMD score was reduced sig- nificantly (all P<0.01), the differences were not significant in comparison between the two groups (all P>0.05). The total effective rate of cognitive improvement was 86.7% (26/30) in the acupuncture group and was 80.0% (24/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05). The total effective rate of the improvement in depression was 93.3% (21/30) in the acupuncture group and was 86.7% (26/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05).

CONCLUSIONAcupuncture could significantly increases CBFV and reduces serum sCD40L expressions in the patients of SIVD complicated with depression, and significantly improves cognitive function and relieves depression symptoms. The efficacy of it is similar to that of western medication. The increase of serum sCD40L expression is possibly involved in the occurrence and development of SIVD. Reducing sCD40L expression contributes to the alleviation of damage induced by cerebral ischemia and reperfusion.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Cerebral Infarction ; complications ; drug therapy ; therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Fluoxetine ; administration & dosage ; Humans ; Male ; Middle Aged ; Nimodipine ; administration & dosage ; Treatment Outcome

In order to determine the suitable harvest time of Dendrobium officinale from different regions in Yunnan province, the drying rate, mannose and glucose peak area ratio, extract, contents of polysaccharide and mannose of D. officinale samples collected from six producing areas in Ynnnan province were determined. The results indicate that drying rate and the contents of polysaccharide and mannose arrived the peak from January to April, extract reached a higher content from September to December, and mannose and glucose peak area ratio from October to February of the coming met the requirment of the Chinese Pharmacopoeia. Hence, the suitable harvesting time of D. officinale in Yunnan province is from December to February of the coming year,according to the experimental results and the request of the Chinese Pharmacopoeia.
China ; Dendrobium ; chemistry ; growth & development ; metabolism ; Glucose ; analysis ; metabolism ; Mannose ; analysis ; metabolism ; Time Factors

OBJECTIVETo explore the effect of Jingui Shenqi pill (JGSQP) with various concentrations at different time points on pituitary adrencorticotropic hormone (ACTH) gene expression level in Shen-Yang deficiency rats.

METHODSThe Shen-Yang deficiency rats were randomly divided into the model control group and the high, medium and low dosage of JGSQP groups. Reverse transcriptase polymerase chain reaction was used to observe the effect of JGSQP on the ACTH mRNA of pituitary tissue in rats treated at different time points (10 d, 20 d, 30 d).

RESULTSAs compared with that in the model group, the ACTH gene expression level was significantly higher in the high dose JGSQP group (P < 0.05), and the increment in the medium dosage group was significantly higher in comparing with that in the high and low dosage groups (P < 0.05 or P < 0.01).

CONCLUSIONThrough up-regulation on ACTH gene expression is possibly one of the mechanisms of JGSQP in treating Shen-Yang deficiency.

Adrenocorticotropic Hormone ; biosynthesis ; genetics ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hypothalamo-Hypophyseal System ; Kidney Diseases ; genetics ; metabolism ; Medicine, Chinese Traditional ; Pituitary Gland ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; genetics ; metabolism

Background Ultraviolet radiation is one of factors of the formation of age-related cataract.Indole-3-carbinol(I3C) is a plant chemical material with inhibitory effect on oxidative-induced cell damage and formation of amyloid fibrils,and the oxidative damage and amyloid fibrils are associated with cataract.However,the relationship between I3C and α-crystallin is in study. Objective This study was to evaluate the effects of ultraviolet-B laser irradiation on the secondary structure of α-crystallin and to explore the protection of I3C to chaperone activity of α-crystallin. Methods The fresh eyeballs were obtained from 1-year-old cattle to prepare the purified lens α-crystallin by gel chromatography.α-Crystallin was isolated from cattle lenses using gel chromatography.The purified α-crystallin was collected using fast protein liquid chromatography ( FPLC ) and exposed to 1:308 nmultraviolet-B at different irradiation intensities ( 23.75,118.75,475.00,1187.50,2375.00,4750.00,11 875.00,23 750.00 mJ/cm2 ) and then to ultraviolet-B 2:308 nm with irradiation intensities of 28 535.00,6730.00,3435.00,1910.00,1040.00 mJ/cm2.Ultraviolet-absorbance spectra,tryptophan fluorescence and N-formylkynurenine (N-FK)fluorescence spectra of both irradiated and non-irradiated α-crystallin were measured.I3C at the concentrations of 50 μmol/L and 100 μmoL/L were added to the α-crystallin solution to perform a catalase (CAT) thermal aggregation to confirm the chaperone activity of the α-crystallin,and the α-crystallin solution without any I3C was used as control.The ratios of A360 between various intervene groups with control group were calculated using spectrophotometry.Results The A280 values of the α-crystallin declined to 10％ at the ultraviolet-B irradiation intensity of 1187.5 mJ/cm2 and that at the intensity of 23.75 J/cm2 lowed to 2％.A negative correlation was seen between the ultraviolet-B irradiation intensity and the A280 value of the α-crystallin (R2=0.925 ) and a positive correlation was found between ultraviolet-B with N-FK ( R2 =0.949 ).Ultraviolet-B irradiation intensity showed a negative correlation with Trp fluorescence intensity (R2 =0.996 ).CAT hot condensed experiment revealed that after addition of different concentrations of indole-3-carbinol,the relative A360 values at various ultraviolet-B irradiation group were significantly higher than those of the control group (P =0.000),and the decreasing degree of chaperone activity of α-crystallin was lower than that of the control group ( P =0.000 ). Conclusions The study suggests that I3C can protect the chaperone activity of α-crystallin from photooxidation,and the ultraviolet-B laser may be a good exposure source compared with ultraviolet lamp.The ultraviolet-B laser irradiation causes the alteration of structure and chaperone activity of α-crystallin.

Adolescent ; Child ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Radius Fractures ; therapy

OBJECTIVETo compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis.

METHODSThirty-six female one-month-old SD rats of (125 +/- 3) g body weight were randomly divided into three groups, 12 rats in each group, one group was orally administered osthole at 9 mg x kg(-1) d(-1), one group was given genistein at 10 mg x kg(-1) d(-1) and another was given equal quantity of distilled water as the control. The body weight was monitored weekly and the bone mineral density (BMD) of total body was measured every month. All rats were sacrificed after three months, the femoral bone mineral density, the serum levels of osteocalcin (OC) and anti-tartaric acid phosphatase 5b (TRACP 5b) were measured by Elisa. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine.

RESULTSNo significant differences were observed between O-treated or GEN group and the control for the food-intake and body weight during three months. However, the rats treated with OST had significant higher BMD for both total body and femur than the control and GEN group. The O-treated rats also had higher level of serum OC and lower level of TRACP 5b. Besides, they owned bigger bone volume/tissue volume, trabecular thickness, trabecular number but smaller trabecular spacing. In the three point bending tests of femurs,they were found to have larger maximum load, the young's modulus and structural model index (SMI).

CONCLUSIONOrally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.

Acid Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Bone Density ; drug effects ; Coumarins ; pharmacology ; Female ; Femur ; diagnostic imaging ; drug effects ; pathology ; Genistein ; pharmacology ; Isoenzymes ; blood ; Osteocalcin ; blood ; Radiography ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

OBJECTIVETo establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field.

METHODSThree 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining.

RESULTSCompared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01).

CONCLUSION4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Chloral Hydrate ; pharmacology ; Cilia ; drug effects ; enzymology ; physiology ; Male ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Radius Fractures ; therapy ; Ulna Fractures ; therapy

OBJECTIVETo investigate the effect of ischemic postconditioning on the expression of rat myocardium matris metalloproteinase-2 (MMP-2) induced by ischemia/reperfusion (I/R) and relationship between its expression and interstitium and the effect on left ventricular function.

METHODSTwenty-four rats were randomly divided into 3 groups (n = 8): sham control (SC) group, ischemic/reperfusion (I/R) group and ischemic postconditioning (IPTC) group. The left ventricular peak systolic pressure and its derivate (+/- dp/dt) were calculated; The amount of myocardium collagenous were determined; The vitality of superoxide dismutase (SOD) and content of malondialdehyde (MDA) of plasma were detected; The activity of myocardium MMP-2 was measured by Western blot and RT-PCR.

RESULTSAs compared with I/R group, IPTC could lower the expression of MMP-2, ameliorate left ventricular function and increase the content of myocardium collagenous. In the meantime, the vitality of superoxide dismutase (SOD) of plasma were greatly enhanced and the content of malondialdehyde (MDA) of plasma were reduced in IFC group.

CONCLUSIONProtective effect of IPIC on myocardium may be due to reduce free radical, lower expression of MMP-2 and protect myocardial interstitium. MMPs plays an important role in the myocardial protection provided by IPTC.

Animals ; Collagen ; metabolism ; Ischemic Postconditioning ; Malondialdehyde ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myocardial Reperfusion Injury ; enzymology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism