Cell Proliferation ; DNA ; chemistry ; Durapatite ; chemistry ; Hep G2 Cells ; Humans ; Liposomes ; chemistry ; Nanoparticles ; chemistry ; Particle Size ; Recombinant Proteins ; chemistry ; Transfection

Background Researches showed that the content of nitric oxide (NO) and nitric oxide synthetase (NOS) increases in blood,aqueous humor and tear of cataract patient.But the function of NO and NOS in cataract formation is still elusive.Objective The aim of this study was to explore the prevention and treatment effect of NOS inhibitor,1-nitro-arginine methyl ester (L-NAME),on galactose cataract.Methods Sixty clean three-week-old Wistar rats were equally and randomly divided into 3 groups.0.9％ Normal saline solution (30 ml/kg) was subcutaneously injected every day for 30 days in the rats of the control group,and 50％ of D-galactose solution (30 ml/kg) was used in the rats of the model and L-NAME group at the same way.L-NAME eye drops was simultaneously administered in the L-NAME group 3 times per day for 30 days.The eyes of the rats were examined under the slit lamp in 10,20 and 30 days,and the degree of lens opacification was scored.Lenses of the rats were obtained at the end of this experiment for the detect of NO,NOS contents.Flow cytometry was used to assay the caspase-3 level of rat lens.Repeated measurement two factor analysis of variance was used to analyze the difference of lens opacification scores in different groups and different time points,and one-way ANOVA was used to analyze the differences of NO,NOS and caspase-3 contents in lens among the groups.Results Lens opacification appeared in 10 days after injection of 50％ D-galactose solution in the rats of the model group and L-NAME group.Lens opacification score was higher among the different groups and different time points (Ftime =435.251,P =0.000 ;Fgroup =395.120,P=0.000).NO content in the lens was (0.45±0.15) μmol/g,(2.67 ± 0.47) μmol/g and (1.68±0.34) μmol/g in the control group,model group and L-NAME group,showing a significant difference (F=58.872,P=0.000).The NOS contents in the lens was (0.0160±0.0020) U/ml,(0.0370±0.0040) U/ml and (0.0270±0.0010) U/ml in the control group,model group and L-NAME group,showing a significant difference (F =66.174,P=0.000).Caspase-3 contents in the lens was (339.4 ± 37.9),(697.7 ± 46.5) and (650.7 ± 53.1),Showing a significant difference among them (F =100.005,P =0.000).Conclusions The increase of NO,NOS and caspase3 levels are associated with lens opacification.Topical administration of L-NAME eye drops can down-regulate NOS content in lens,reduce the NO formation and inhibit the apoptosis of lens epithelial cells.

Adult ; Bronchial Hyperreactivity ; physiopathology ; Bronchial Provocation Tests ; Chronic Disease ; Cough ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Respiratory Function Tests

OBJECTIVETo explore effects of Shenqi preparation,Traditional Chinese Medicine, on anti-fatigue and anti-oxidant functions.

METHODSOne hundred and twenty mice were randomly divided into control group and 3 experimental groups. The high, medium and low-dose of Shenqi preparation were given to the 3 experimental groups respectively, while distilled water to the control group for 15 d. The loaded swimming time, the level of lactate, serum urea nitrogen (SUN), muscle and liver glycogen, liver super-oxide dismutase (SOD), the content of malondialdehyde (MDA), glutathione peroxidase(GSH-Px) were assayed.

RESULTSThe loaded swimming test showed that the exhausted swimming time of 3 experimental groups [(296.0 +/- 25.3)s, (437.0 ĝ 38.9)s, (595.0 +/- 53.9)s respectively] was longer than that of control group [(231.0 +/- 22.5)s, P < 0.05, P < 0.01]. The liver glycogen content of the high and medium-dose experimental groups were higher than that of control group respectively (P < 0.01). The SUN content of each experimental group was less than that of the control group (P < 0.01, P < 0.05). Moreover,in the medium and high dose experimental groups, less accumulation of lactate was found (P < 0.01, P < 0.05), and the content of liver SOD and GSH-Px was higher (P < 0.01, P < 0.05). The content of liver MDA in high-dose experimental group was less than that of the control group (P < 0.05).

CONCLUSIONShenqi preparation, especially the high and medium-dose experimental groups, is able to improve exercise tolerance and has anti-fatigue and anti-oxidant effects in mice.

Animals ; Antioxidants ; metabolism ; Blood Urea Nitrogen ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Fatigue ; drug therapy ; metabolism ; Glutathione Peroxidase ; metabolism ; Glycogen ; metabolism ; Lactic Acid ; blood ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Physical Conditioning, Animal ; Superoxide Dismutase ; metabolism

OBJECTIVETo explore the intervening and therapeutic effect of Cordyceps mycelia extract (CME) on liver cirrhosis induced by dimethylnitrosamine (DMN) in rats.

METHODSRat liver cirrhosis model was established by peritoneal injection of DMN at a dose of 10 microg/kg, once daily in the first 3 days of every week for 4 successive weeks. Experimental study on CME-intervention was conducted from the beginning of modeling to the end of the 4th week, while the CME-treatment experiment was carried out from the 4th week of modeling, when terminating the modeling factor, to the end of the 8th week, by administering CME at a dose of 0. 74 g/( kg d) once a day. Animals were killed in batches on the 3rd day, the 2nd (T1), 4th (T2), 6th (T3) and 8th (T4) week after modeling, to observe the histopathologic change in liver and the immunohistochemical staining of alpha-smooth muscle actin (alpha-SMA) and collagen type I (Col I), determine the content of hydroxyproline (Hyp) in liver, and the liver function was tested as well.

RESULTSCME-intervention experiment showed that as compared to those in the modeled rats at corresponding time points, in rats at T1 and T2, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity and total bilirubin (TBIL) content were significantly lower, and albumin (Alb) obviously higher; while at T2, Hyp content, ct-SMA and Col I positive expression were significantly lower (P < 0.05), the proliferation of collagen fibre attenuated. CME-treatment experiment showed that as compared to those in the modeled rats at corresponding time points, lower serum ALT, AST activity and TBIL content, and higher serum level of Alb were shown in rats at T1; and lower Hyp content, liver collagen fibre, and alpha-SMA positive expression were shown at T1 and T2; while less Col I positive expression at T2 was also shown in them (all P < 0.05).

CONCLUSIONCME could not only prevent the development of liver cirrhosis induced by DMN in rats, but also effectively promote the reversion of already formed liver cirrhosis, having a favourable prospect of clinical application.

Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Biological Factors ; therapeutic use ; Cordyceps ; chemistry ; Dimethylnitrosamine ; adverse effects ; Humans ; Liver ; metabolism ; Liver Cirrhosis ; chemically induced ; drug therapy ; enzymology ; genetics ; Male ; Moths ; chemistry ; Mycelium ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Treatment Outcome

Objective To know the antitumor effect and value of vinorelbine in chemotherapy with relapsed breast cancer.Methods To receive breast cancer tissue from primary and relapsed patients by surgery,then to obtain mammary cancer cells by collagenase.The antitumor effect of 5 chemotherapy drugs with breast cancer cells was measured by culture in vitro and MTT.Results With primary cancer cells from primary breast cancer,the antitumor effect of paclitaxel and docetaxel(sensitivity is 91.04%,92.54% respec- tively) is better than adriamycin,epirubicin and gemcitabine(sensitivity is 73.13%,74.63%,71.64% re- spectively;P

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P＜0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P＜0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P＜0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P＜0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P＜0.01) and in the gastric cancer tissues (P＜0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P＜0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P＜0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.

Objective:To observe the effect of moxibustion on T lymphocyte subsets in peripheral blood of rats with gastric cancer. Methods:Sixty healthy Sprague-Dawley (SD) rats were adaptively fed for one week. By the random number table method, 10 rats were randomly selected as a blank group, and 12 rats were randomly selected to simulate the tumor transplantation process; after transplantation, 10 rats were randomly selected as a sham operation group. The remaining 38 rats were used to prepare gastric cancer models by gastric transplantation of the Walker-256 tumor tissue; 8 rats were randomly selected to verify the successful modeling after 7 d; the remaining 30 rats were randomly divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. From the first day of enrollment, the rats in the moxibustion group received mild moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36) (the first group) and bilateral Pishu (BL 20) and Weishu (BL 21) (the second group), and the two groups of acupoints were alternated every other day. The rats in the infrared group received infrared radiation on the stomach area and the area on the back between the T12-T13 spinous processes, the two areas were alternated every other day. Rats in the moxibustion group and the infrared group were treated for 20 min each time, once a day for 21 d. Rats in the blank group, the sham operation group, and the model group were simultaneously grasped and fixed, and no other treatment was performed. After 21 d of intervention, the rats in each group were fasted for 12 h, and blood was collected from the orbits. The numbers of CD3+, CD3+CD4+, CD3+CD8+ T lymphocytes in peripheral blood were determined by flow cytometry, and the ratio of CD3+CD4+/CD3+CD8+ was calculated. The rats were sacrificed and the thymus was dissected under sterile conditions to calculate the thymus index. Results:Compared with the blank group, the thymus index, peripheral blood CD3+, CD3+CD4+, CD3+CD8+ and CD3+CD4+/CD3+CD8+ ratio in the sham operation group did not change significantly (allP>0.05). Compared with the blank group, the thymus index of the model group was increased (P<0.05), the CD3+ and CD3+CD8+ T lymphocytes were increased (bothP<0.01), and the CD3+CD4+/CD3+CD8+ ratio was decreased (P<0.05). Compared with the model group, the thymus index of the moxibustion group was increased (P<0.01), and CD3+, CD3+CD4+ and CD3+CD8+ T lymphocytes and the ratio of CD3+CD4+/CD3+CD8+ in peripheral blood were increased (allP<0.05). Compared with the infrared group, the thymus index of the moxibustion group was significantly increased (P<0.05), the CD3+ and CD3+CD4+ T lymphocytes in the peripheral blood were significantly increased (bothP<0.01), and the CD3+CD8+ was increased (P<0.05). Conclusion:Moxibustion can significantly increase the thymus index of gastric cancer-bearing rats and activate CD3+CD4+ and CD3+CD8+ T lymphocytes in peripheral blood.

OBJECTIVETo investigate the effects of celecoxib combined with radiotherapy on apoptosis of CNE-2Z cell lines and the potential mechanisms.

METHODSFour groups were used, a control, celecoxib (25 micromol/L celecoxib), irradiation (8 Gy X ray) and celecoxib plus irradiation. The radiosensitising effect was detected by clone formation experiment. Flow cytometry was used to detect the apoptosis rate of cells. The expressions of Bcl-2 and Bax were assessed by immunocytochemistry. Western blot was used to examine the expression of Caspase-3.

RESULTSCelecoxib enhanced the radiosensitivity of CNE-2Z cells. In experimental group, the mean surviving fraction and the mean lethal dose of CNE-2Z cells were 0.50 and 2.36 respectively. Compared with the irradiated group, there was significant differences between the two groups (P < 0.01). Celecoxib combined with radiotherapy up-regulation the expression of Bax. The score of the expression of Bax in the control group and the experimental group were 1.221 +/- 0.116 and 2.758 +/- 0.256 respectively. Celecoxib combined with radiotherapy could inhibit the expression of the protein of Bcl-2. The score of the expression of Bcl-2 in the control group and the experimental group were 2.559 +/- 0.144 and 1.253 +/- 0.114 respectively, with significant differences (P < 0.01). Celecoxib combined with radiotherapy could increase the apoptosis rate of tumor cells with significant differences (F = 7.63, P < 0.01). Western blot showed that the expression of Caspase-3 was strengthened.

CONCLUSIONCelecoxib combined with radiotherapy could induce apoptosis and enhance the radiosensitivity of human nasopharyngeal carcinoma CNE-2Z cell lines.

Apoptosis ; drug effects ; radiation effects ; Carcinoma ; Caspase 3 ; metabolism ; Celecoxib ; Cell Line, Tumor ; drug effects ; radiation effects ; Humans ; Nasopharyngeal Neoplasms ; pathology ; therapy ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrazoles ; pharmacology ; Radiotherapy ; Sulfonamides ; pharmacology ; bcl-2-Associated X Protein ; metabolism

Objective To evaluate the effect of PTH gene polymorphisms on bone mineral density (BMD) at multiple skeletal sites in normal females.Methods PTH gene phenotype was determined by PCR-RFLP of restriction enzyme Bst BⅠin 596 females aged (46.3?13.7) years (20-80 years),and PCR products with or without enzymolytic site were considered as genotype B or genotype b respectively.BMDs of the anteropesterior spine (AP) and supine lateral spine (Lat) of lumbar vertebrae (L_1-L_4),femoral neck (FN),total hip (T-hip), Ward's triangle (Ward),Trochanter (Troch),forearm [radius+ulna ultradistal (RUUD) and total area of radius + ulna (RUT) ] were measured by DEXA (QDR4500A).Results (1) Hardy-Weinberg equilibrium was evident for PTH polymorphisms.The frequencies of genotype were BB 0.784,Bb 0.208,bb 0.008 and frequencies of alleles B,b were 0.888 and 0.112 respectively in 596 normal females.Frequencies of BB,Bb,bb genotypes were 0.781,0.210,and 0.009 respectively in 347 postmenopausal women and their frequencies of alleles B,b were 0.886,0.114.No significant difference was found between post- and premenopausal women in genotype frequen- cy.(2) Comparing their BMDs of AP,Lat,FN,T-hip,Ward,Troch,RUUD and RUT,there was no significant difference between BB and Bb genotypes of pre- and postmenopansal women groups.(3) Logistic regression analysis failed to show any statistical difference between normal and osteoporosis women with regard to PTH phenotype.Conclusion PTH gene polymorphism has little effect on BMD in normal females.